Offspring Of Pregnant Females Research Articles

Modifications of Behavior and Inflammation in Mice Following Transplant with Fecal Microbiota from Children with Autism

Autism spectrum disorder (ASD) is a neurodevelopmental disorder displaying the modification of complex human behaviors, characterized by social interaction impairments, stereotypical/repetitive activities and emotional dysregulation. In this study, fecal microbiota transplant (FMT) via gavage from autistic children donors to mice, led to the colonization of ASD-like microbiota and autistic behaviors compared to the offspring of pregnant females exposed to valproic acid (VPA). Such variations seemed to be tightly associated with increased populations of Tenericutes plus a notable reduction (p < 0.001) of Actinobacteria and Candidatus S. in the gastrointestinal region of FMT mice as compared to controls. Indeed altered behaviors of FMT mice was reported when evaluated in the different maze tests (light dark, novel object, three chamber tests, novel cage test). Contextually, FMT accounted for elevated expression levels of the pro-inflammatory factors IL-1β, IL-6, COX-1 and TNF-α in both brain and small intestine. Villous atrophy and inflammatory infiltration (Caspase 3 and Ki67) were increased in the small intestine of FMT and VPA mice compared to controls. Moreover, the observed FMT-dependent alterations were linked to a decrease in the methylation status. Overall, findings of the present study corroborate a key role of gut microbiota in ASD. However, further investigations are required before any possible manipulation of gut bacteria with appropriate diets or probiotics can be conducted in ASD individuals.

Increased alpha and beta cell mass during mouse pregnancy is not dependent on transdifferentiation.

Maternal pancreatic beta-cell mass (BCM) increases during pregnancy to compensate for relative insulin resistance. If BCM expansion is suboptimal, gestational diabetes mellitus can develop. Alpha-cell mass (ACM) also changes during pregnancy, but there is a lack of information about α-cell plasticity in pregnancy and whether α- to β-cell transdifferentiation can occur. To investigate this, we used a mouse model of gestational glucose intolerance induced by feeding low-protein (LP) diet from conception until weaning and compared pregnant female offspring to control diet-fed animals. Control and LP pancreata were collected for immunohistochemical analysis and serum glucagon levels were measured. In order to lineage trace α- to β-cell conversion, we utilized transgenic mice expressing yellow fluorescent protein behind the proglucagon gene promoter (Gcg-Cre/YFP) and collected pancreata for histology at various gestational timepoints. Alpha-cell proliferation increased significantly at gestational day (GD) 9.5 in control pregnancies resulting in an increased ACM at GD18.5, and this was significantly reduced in LP animals. Despite these changes, serum glucagon was higher in LP mice at GD18.5. Pregnant Gcg-Cre/YFP mice showed no increase in the abundance of insulin+YFP+glucagon- cells (phenotypic β-cells). A second population of insulin+YFP+glucagon+ cells was identified which also did not alter during pregnancy. However, there was an altered anatomical distribution within islets with fewer insulin+YFP+glucagon- cells but more insulin+YFP+glucagon+ cells being present in the islet mantle at GD18.5. These findings demonstrate that dynamic changes in ACM occur during normal pregnancy and were altered in glucose-intolerant pregnancies.

Effects of persistent environmental pollutants on the HPA-axis

Background Many persistent chemicals which are abundant in nature accumulate through food webs and are transferred to the offspring of pregnant females through the placenta and mother’s milk. Some of these chemicals can adversely affect neurodevelopment, the immune system or act as endocrine disruptors. Effects of persistent organic pollutants (POPs) on the hypothalamus-pituitary-adrenal (HPA) axis have been relatively poorly studied in comparison with other endocrine axes. However, blood and tissue levels of certain POPs have been shown to correlate with cortisol levels in polar bears and to affect adrenal function in fish and other wildlife [1]. The HPA axis and its end product, cortisol, is not only crucial for adequate responses to stress, it is also involved in normal foetal development, as well as regulation of metabolism, immune function, blood pressure and other bodily functions. The aim of our research has been to investigate endocrine disrupting effects of different polychlorinated biphenyls (PCBs) and mixtures of POPs. Effects were observed on the adrenal steroidogenesis and foetal development of the adrenal cortex. Special attention was given to the synthesis of cortisol.

Increased Myogenic Responses in Uterine but not Mesenteric Arteries from Pregnant Offspring of Diet-Restricted Rat Dams1

Results of epidemiological and animal studies suggest a link between poor in utero growth and cardiovascular disease in adult offspring. Few studies, however, have examined the effects of maternal undernutrition on the vasculature of pregnant female offspring, and to our knowledge, no studies have examined myogenic responses, which are essential to vascular tone development, in these animal models. Thus, myogenic responses were assessed in radial uterine arteries of pregnant female offspring to determine if diet restriction during pregnancy could contribute to transgenerational effects. These results were compared to those in mesenteric arteries, which greatly contribute to peripheral vascular resistance. Myogenic responses in the presence and absence of inhibitors for nitric oxide synthase (NOS) and prostaglandin H synthase (PGHS) were measured in arteries isolated from pregnant, 3-mo-old female offspring of control-fed (C(off)) and globally diet-restricted (DR(off)) rat dams. Although no differences were found in pregnancy weight gain, litter size, or fetal weights, placental size was significantly reduced in DR(off) compared to C(off). Enhanced myogenic reactivity was observed at the highest pressure tested (110 mm Hg) in uterine, but not in mesenteric, arteries from DR(off) compared to C(off). Inhibition of NOS, but not of PGHS, significantly increased myogenic responses in uterine arteries at pressures greater than 80 mm Hg in C(off) but, interestingly, not in DR(off) compared to untreated uterine arteries. Thus, impaired uterine vascular function in diet-restricted pregnant rat dams, which leads to similar impairment in their pregnant offspring, may be a mechanism through which transgenerational effects of unhealthy pregnancies occur.

Protein-deficient diet in pregnant females impairs functional activity of enzyme systems in digestive and non-digestive organs in the offspring.

Activity of disaccharidases in various portions of the small intestine markedly increased in 6-month-old offspring of pregnant females feeding a protein-deficient diet. Activities of amino- and dipeptidases decreased in the kidneys, while alkaline phosphatase activity remained practically unchanged. Probably, information about protein restriction in nutrition of pregnant females is stored in the biochemical memory of adult offspring and promotes the development of metabolic disturbances.

Maternal protein restriction in the rat impairs resistance artery but not conduit artery function in pregnant offspring

Dietary protein restriction during gestation has been shown to produce vascular dysfunction in pregnant rats and hypertension in their offspring. However, no studies have to date examined the effects of such 'programming' on the vascular function of female offspring when they in turn become pregnant. We have therefore studied isolated conduit and resistance artery function from pregnant female offspring of control (C, 18 % casein) and protein-restricted (PR, 9 % casein) pregnant dams. There were no differences in birth weight, weight gain during pregnancy, litter size, fetal weight, placental weight, fetal : placental weight ratio or organ weights between the C and PR groups. In isolated mesenteric arteries, the vasodilatation in response to the endothelial-dependent vasodilator acetylcholine (ACh) and the beta-adrenoceptor agonist isoprenaline was decreased in the PR group, while there were no differences in the constriction in response to potassium (125 mM) or the alpha1-adrenoceptor agonist phenylephrine (PE). No differences in any responses were seen in the isolated thoracic aorta. We conclude that dietary protein restriction in pregnancy programmes vasodilator dysfunction in isolated resistance arteries of female offspring when they become pregnant, but does not affect conduit arteries.

Germ Cell Regulation of Sertoli Cell Transferrin mRNA Levels

Sertoli cells cultured in the presence of germ cells responded by increasing the level of transferrin mRNA 3-fold as determined by solution hybridization and Northern blot analysis. In contrast, the steady state levels of other mRNAs, including sulfated glycoprotein 1 (SGP-1), sulfated glycoprotein 2 (SGP-2), transferrin receptor, regulatory subunit of cAMP dependent protein kinase, and ferritin light chain, were not influenced by coculture with germ cells. The transferrin mRNA stimulatory activity was found in conditioned medium from germ cells but was not associated with germ cell membrane components. The activity was abolished by treatment of the medium with trypsin. Partial characterization and isolation of the protein(s) from conditioned medium indicated that it has an apparent mol wt between 10 and 30 K. Studies using inhibitors of protein and nucleic acid synthesis indicated that the stimulation of transferrin mRNA by germ cell conditioned medium required both transcription and translation. Sertoli cell enriched (germ cell depleted) testes were obtained from male offspring of pregnant females irradiated at the 19th day of gestation. Testicular transferrin mRNA levels from irradiated rats decreased in comparison to levels in the normal rat, whereas SGP-2 mRNA levels were unchanged. These studies demonstrate that germ cell secretions may interact with Sertoli cells to specifically increase the level of transferrin mRNA and that this interaction may be a mechanism by which germ cells regulate the flow of iron across the seminiferous epithelium.

Neurogenetic patterns in the medial limbic cortex of the rat related to anatomical connections with the thalamus and striatum

[ 3H]Thymidine autoradiography was used to investigate neurogenesis in all areas of the limbic cortex in the medial wall of the hemisphere. The experimental animals were the offspring of pregnant females injected with [ 3H]thymidine on 2 consecutive days: Embryonic Day (E)13–E14, E14–E15, …E21–E22, respectively. On Postnatal Day (P)60, the proportion of neurons originating during 24-h periods were quantified at nine anteroposterior levels. Three types of neurogenetic gradients are found. (i) Deep cells are older than superficial cells: layer VI is generated mainly on E15–E16, layer V on E16–E18, and layers IV-II on E18–E20. (ii) There is a ventral/older to dorsal/younger gradient between the dorsal peduncular, infralimbic, and anterior cingulate areas rostral to the genu of the corpus callosum. A ventral/older to dorsal/younger gradient is also found between superficial cells (layers II–IV) in anterior cingulate (CG3/CG1), posterior cingulate (CG2/CG1), and retrosplenial areas (RSG/RSA). (iii) An anterior/older to posterior/younger gradient is found between areas throughout the medial limbic cortex. Some of these neurogenetic patterns correlate with anatomical interconnections between the supracallosal medial limbic cortex (posterior cingulate and retrosplenial areas) and the anteroventral/anteromedial thalamic nuclei: older thalamic cells have longer axons that terminate in cortical areas containing younger cells, while younger thalamic cells have shorter axons that terminate in cortical areas containing older cells. Projections from the medial limbic cortex to the striatum also correlate with neurogenetic gradients: older cortical source cells in the infralimbic area project to the older striatal cells in the enkephalin-rich patches, while younger cortical source cells in the cingulate areas project to younger striatal cells in the surrounding matrix.

Development of the lateral and medial limbic cortices in the rat in relation to cortical phylogeny

[ 3H]Thymidine autoradiography was used to investigate neurogenesis of the lateral limbic cortex and morphogenesis of the medial and lateral limbic cortices in adult and embryonic rat brains. Ontogenetic patterns in the limbic cortex are unique because some neurogenetic gradients are linked to those in neocortex, others are linked to those in paleocortex. These findings are related to hypotheses of cortical phylogeny. The experimental animals used for neurogenesis were the off-spring of pregnant females injected with [ 3H]thymidine on 2 consecutive days: Embryonic Day (E)13–E14, E14–E15, …E21–E22, respectively. On Postnatal Day (P)60, the proportion of neurons originating during 24-h periods were quantified at nine anteroposterior levels and one sagittal level. Similar to neocortex, deep cells are generated earlier than superficial cells throughout the lateral limbic cortex: layer VI mainly on E14–15, layer V on E15–E16, and layers IV-II on E16–E18. There is a ventral/older to dorsal/younger neurogenetic gradient between the ventral agranular insular, dorsal agranular insular, and gustatory cortical areas and between ventral and dorsal orbital areas beneath the frontal pole. Similar to paleocortex below the rhinal sulcus, limbic cortex in the rhinal sulcus has a “sandwich” gradient: the older posterior agranular insular area is sandwiched by anterior and posterior younger areas (ventral agranular insular and perirhinal). To study morphogenesis, pregnant females were given single injections of [ 3H]thymidine during gestation and embryos were removed in successive 24-h intervals (sequential-survival). Neurogenesis finishes first in ventral limbic areas, later in dorsal limbic areas, and latest in neocortical areas. The cortical plate in the region of the medial and lateral limbic cortices does not have a separate subplate layer as is found in the region of the neocortex. Instead, layer VI in the limbic cortices has unusually older cells that are generated simultaneously with subplate cells.

Development of layer I and the subplate in the rat neocortex

Development of layer I and the subplate of the rat neocortex was examined with [ 3H]thymidine autoradiography. The experimental animals used for neurogenesis were the offspring of pregnant females injected with [ 3H]thymidine on 2 consecutive days: Embryonic Day (E) 13–E14, E14–E15, … E21–E22, respectively. On Postnatal Day 5, the proportion of layer I and subplate cells originating during 24-h periods were quantified at three anteroposterior levels. Presumptive Cajal-Retzius cells (large horizontal cells) are generated mainly on E14 and subplate cells on E14 and E15 (“outside-in” gradient). Both populations are generated earlier than cells in the cortical plate, which has an “inside-out” gradient. The subplate also has a ventrolateral/older to dorsomedial/younger neurogenetic gradient. The small- to medium-sized horizontal cells in layer I have an extensive period of neurogenesis with an “outside-in” gradient. To study morphogenesis, pregnant females were given single injections of [ 3H]thymidine during gestation and embryos were removed in successive 24-h intervals (sequential-survival). On E15 and E16, cells accumulate outside the neuroepithelium in the primordial plexiform layer with older presumptive Cajal-Retzius cells superficial and younger presumptive subplate cells deep. The Cajal-Retzius cells permanently settle superficially among a first system of extracellular channels that appears on E14. Before reaching their final settling sites, subplate cells form the incipient cortical plate in the ventrolateral neocortex on E16. On E17, a second system of extracellular channels appears below the cortical plate. On E18 and E19, subplate cells leave the cortical plate and permanently settle among the deep extracellular channels in a separate layer.

Neurogenetic and morphogenetic heterogeneity in the bed nucleus of the stria terminalis.

Neurogenesis and morphogenesis in the rat bed nucleus of the stria terminalis (strial bed nucleus) were examined with [3H]thymidine autoradiography. For neurogenesis, the experimental animals were the offspring of pregnant females given an injection of [3H]thymidine on 2 consecutive gestational days. Nine groups of embryos were exposed to [3H]thymidine on E13-E14, E14-E15,... E21-E22, respectively. On P60, the percentage of labeled cells and the proportion of cells originating during 24-hour periods were quantified at six anteroposterior levels in the strial bed nucleus. On the basis of neurogenetic gradients, the strial bed nucleus was divided into anterior and posterior parts. The anterior strial bed nucleus shows a caudal (older) to rostral (younger) neurogenetic gradient. Cells in the vicinity of the anterior commissural decussation are generated mainly between E13 and E16, cells just posterior to the nucleus accumbens mainly between E15 and E17. Within each rostrocaudal level, neurons originate in combined dorsal to ventral and medial to lateral neurogenetic gradients so that the oldest cells are located ventromedially and the youngest cells dorsolaterally. The most caudal level has some small neurons adjacent to the internal capsule that originate between E17 and E20. In the posterior strial bed nucleus, neurons extend ventromedially into the posterior preoptic area. Cells are generated simultaneously along the rostrocaudal plane in a modified lateral (older) to medial (younger) neurogenetic gradient. Ventrolateral neurons originate mainly between E13 and E16, dorsolateral neurons mainly between E15 and E16, and medial neurons mainly between E15 and E17. The youngest neurons are clumped into a medial "core" area just ventral to the fornix. For morphogenesis, pregnant females were given a single injection of [3H]thymidine during gestation, and their embryos were removed either 2 hours later (short survival) or in successive 24-hour periods (sequential survival). The embryonic brains were examined to locate areas of intensely labeled cells in the putative neuroepithelium of the strial bed nucleus, to trace migratory waves of young neurons, and to establish their final settling locations. Two different neuroepithelial sources produce neurons for the strial bed nucleus. The anterior strial bed nucleus is generated by a neuroepithelial zone at the base of the inferior horn of the lateral ventricle from the anterior commissural decussation area forward to the primordium of the nucleus accumbens.(ABSTRACT TRUNCATED AT 400 WORDS)

Development of the preoptic area: Time and site of origin, migratory routes, and settling patterns of its neurons

Neurogenesis and morphogenesis in the rat preoptic area were examined with [3H]thymidine autoradiography. For neurogenesis, the experimental animals were the offspring of pregnant females given an injection of [3H]thymidine on two consecutive gestational days. Nine groups were exposed to [3H]thymidine on embryonic days E13-E14, E14-E15, E21-E22, respectively. On postnatal day P5, the percentage of labeled cells and the proportion of cells originating during 24-hr periods were quantified at four anteroposterior levels in the preoptic area. Throughout most of the preoptic area there is a lateral to medial neurogenetic gradient. Neurons originate between E12-E15 in the lateral preoptic area, between E13-E16 in the medial preoptic area, between E14-E17 in the medial preoptic nucleus, and between E15-E18 in the periventricular nucleus. These structures also have intrinsic dorsal to ventral neurogenetic gradients. There are two atypical structures: (1) the sexually dimorphic nucleus originates exceptionally late (E15-E19) and is located more lateral to the ventricle than older neurons; (2) in the median preoptic nucleus, where older neurons (E13-E14) are located closer to the third ventricle than younger neurons (E14-E17). For an autoradiographic study of morphogenesis, pregnant females were given a single injection of [3H]thymidine during gestation, and their embryos were removed either two hrs later (short survival) or in successive 24-hr periods (sequential survival). Short-survival autoradiography was used to locate the putative neuroepithelial sources of preoptic nuclei, and sequential survival autoradiography was used to trace the migratory waves of young neurons and their final settling locations. The preoptic neuroepithelium is located anterior to and in the front wall of the optic recess. The neuroepithelium lining the third ventricle is postulated to contain a mosaic of spatiotemporally defined neuroepithelial zones, each containing precursor cells for a specific structure. The neuroepithelial zones and the migratory waves originating from them are illustrated. Throughout most of the preoptic area, neurons migrate predominantly laterally. The older neurons in the lateral preoptic area migrate earlier and settle adjacent to the telencephalon. Younger neurons migrate in successively later waves and accumulate medially. The sexually dimorphic neurons are exceptional since they migrate past older cells to settle in the core of the medial preoptic nucleus.(ABSTRACT TRUNCATED AT 400 WORDS)

Neurogenesis in the anterior olfactory nucleus and its associated transition areas in the rat brain

Neurogenesis in the rat olfactory peduncle was examined with [3H]thymidine autoradiography. Animals in the prenatal groups were the offspring of pregnant females given an injection of [3H]thymidine on two consecutive gestation days. Nine groups of embryos were exposed to [3H]thymidine on embryonic days (E) E13–E14, E14–E15, … E21–E22, respectively. One group of postnatal animals was given four consecutive injections of [3H]thymidine on postnatal days (P) P0-P3. On P60, the percentage of labeled cells and the proportion of cells originating during either 24 or 48 hr periods were quantified at seven anatomical levels through both the anterior olfactory nucleus and the transition areas. A caudal (older) to rostral (younger) neurogenetic gradient is found both within and between structures in the olfactory peduncle. Neurons in the dorsal, lateral, and ventral-lateral transition areas are generated mainly between E14 and E19, those in the anterior olfactory nucleus mainly between E15 and E21. Only 3–4% of the neurons in the most anterior pars lateralis and pars dorsalis originate after birth. All parts of the anterior olfactory nucleus show a strong superficial (older) to deep (younger) neurogenetic gradient (the “outside-in” pattern). In contrast, neurons in the ventral—lateral transition area and in the dorsal transition area originate in a deep to superficial neurogenetic gradient (the “inside-out” pattern), suggesting that these areas are, in reality, primary olfactory cortex. The lateral transition area is truly “transitional”, showing no neurogenetic gradient along the superficial-deep plane. The medial transition area originates between E15 and E19 in a center (older) to edge (younger) “sandwich” neurogenetic gradient along the rostrocaudal plane, a pattern apparently unrelated to neurogenetic gradients in other olfactory peduncle structures. These data suggest that characteristic patterns of neurogenesis, namely the “insideout” vs the “outside-in” gradients, permit the assignment of different structures to cortical vs ganglionic cytoarchitectonic components of the olfactory relay system.

Neurogenesis in the rat primary olfactory cortex

Neurogenesis in the rat primary olfactory cortex was examined with [3H]thymidine autoradiography. The experimental animals were the offspring of pregnant females given an injection of [3H]thymidine on two consecutive gestation days. Nine groups of embryos were exposed to [3H] thymidine on E13–E14, E14–E15, … E21–E22, respectively. On P60, the percentage of labeled cells and the proportion of cells originating during 24 hr periods were quantified at selected anatomical levels of the anterior and posterior piriform cortex, dorsal lateral peduncular cortex, and posterior two-thirds of the ventral agranular insular cortex. Throughout most of the primary olfactory cortex, deep cells are generated earlier than superficial cells: the ‘dinside-out’ pattern. Neurons in the anterior (prepiriform) cortex are located lateral to the caudal anterior olfactory nucleus and olfactory tubercle, and are generated mainly between E14 and E18 in a caudal (older) to rostral (younger) neurogenetic gradient. Neurons in the posterior (periamygdaloid) cortex are located lateral to the caudal olfactory tubercle and amygdala, and are generated mainly between E14 and E17 simultaneously along the rostrocaudal plane. Superficial cells in the piriform cortex have some additional neurogenetic gradients; ventromedial cells forming transition zones with either the olfactory tubercle or amygdala originate earlier than cells located dorsally and laterally. In the posterior piriform cortex, younger neurons are located at middle dorsoventral levels while older neurons lie above and below. Neurons in the dorsolateral peduncular cortex originate between E14 and E20 in a caudal to rostral gradient of neurogenesis; caudal parts also have a lateral to medial neurogenetic gradient. The most lateral part of the dorsolateral peduncular cortex is unique and does not have the typical ‘inside-out’ cortical neurogenetic gradient. Neurons in the ventral agranular insular cortex (area 13) originate mainly between E15 and E17 in combined caudal to rostral and ventral to dorsal neurogenetic gradients. The neurogenetic gradients in the primary olfactory cortex, along with patterns of neurogenesis throughout the olfactory projection field are related to the termination patterns of afferents from the main olfactory bulb.

Neurogenesis in the olfactory tubercle and islands of Calleja in the rat

Neurogenesis in the rat olfactory tubercle and islands of Calleja was examined with [3H]thymidine autoradiography. Animals in the prenatal groups were the offspring of pregnant females given an injection of [3H]thymidine on two consecutive gestational days. Ten groups of embryos (E) were exposed to [3H]thymidine on E12–E13, E13–E14 4 E21–E22, respectively. Three groups of postnatal animals (P) were given four consecutive injections of [3H]thymidine on P0–P3, P2–P5, and P4–P7, respectively. On P60, the percentage of labeled cells and the proportion of cells originating during either 24 or 48 h periods were quantified at several anatomical levels. Three populations of neurons were studied: 1.(1) large cells in layer III2.(2) small to medium-sized cells in layers II and III, and in the striatal bridges3.(3) granule cells in the islands of Calleja. Neurogenesis is sequential between these three populations with No. 1 oldest and No. 3 youngest. The large neurons in layer III originate mainly between E13 and E16 in a strong lateral-to-medial gradient. Neurons in population No. 2 are generated between E15 and E20, also in a lateral-to-medial gradient; neurogenesis is simultaneous along the superficialdeep plane. Granule cells in the smaller islands of Calleja are generated between E17 and E22 in combined lateral-to-medial and superficial-to-deep gradients. Neurons in the large island of Calleja are generated mainly between E19 and E22 in a strong rostral-to-caudal gradient. Neurogenesis is reduced to < 10% in both populations No. 2 and No. 3, occurring mainly between P0 and P4. The neurogenetic patterns in populations No. 2 and No. 3 are similar to those in the striatum, while neurogenesis in population No. 1 fits into the pattern of the globus pallidus and substantia innominata. These developmental patterns indicate that the olfactory tubercle is a mixed striato-pallidal system rather than an olfactory cortical area. The lateral-to-medial neurogenetic gradient shown in each neuronal population in the olfactory tubercle correlates with both differential anatomical projections and differential neurochemical characteristics along the lateral-medial plane.

Neurogenesis of the magnocellular basal telencephalic nuclei in the rat

Neurogenesis in the magnocellular basal telencephalic nuclei of the rat was examined with [3H]thymidine autoradiography. The experimental animals were the offspring of pregnant females given two injections of [3H]thymidine on consecutive embryonic (E) days (E12–E13, E13–E14, … E21–E22). On postnatal day (P) 60, the percentage of labeled cells and the proportion of cells originating during 24 h periods were quantified at several anatomical levels throughout the magnocellular basal telencephalic nuclei. The neurons of the horizontal limb of the diagonal band originate mainly between E13 and E16 in a combined rostral-to-caudal and lateral-to-medial gradient. The neurogenetic gradients in the horizontal limb are continued by generation patterns of cells in the vertical limb of the diagonal band-medial septal complex, the large cells in the polymorph layer of the olfactory tubercle, and the large cells of the anterior amygdaloid area. The substantia innominata originates between E13 and E17 in combined caudal-to-rostral and lateral-to-medial gradients. The globus pallidus originates between E13 and E17 in combined caudal-to-rostral, ventral-to-dorsal and medial-to-lateral gradients. The entopeduncular nucleus originates between E12 and E14 in a ‘sandwich’ gradient where neurons in the core of the nucleus are older than those in either the anterior or posterior ends. There is an overall superficial (ventral) to deep (dorsal) neurogenetic gradient between the magnocellular basal nuclei present at any given rostrocaudal level. An important finding is that neurogenetic gradients in the individual components of the magnocellular basal nuclei are alike (with the possible exception of the entopeduncular nucleus) indicating they are part of a single system. Finally, evidence is presented that neurogenetic gradients in the magnocellular basal telencephalic neurons can be correlated with their anatomical projections to the cerebral cortex.

Neurogenesis in the rat neostriatum

Neurogenesis in the rat neostriatum was examined with [3H]thymidine autoradiography. For the animals in the prenatal groups, the initial [3H]thymidine exposures were separated by 24 h; they were the offspring of pregnant females given two injections on consecutive embryonic (E) days (E13–E14, E14–E15, … E21–E22). For the animals in the postnatal (P) groups, the initial [3H]thymidine exposures were separated by 48 h, each group receiving four consecutive injections (P0–P3, P2–P5, P4–P7). On P60, the percentage of labeled cells and the proportion of cells originating during either 24 or 48 h periods were quantified at several anatomical levels for both the large and medium-sized neurons. Neurogenesis of the large neurons occurs mainly between E13 and E16 in a strong caudal-to-rostral gradient. The medium-sized neurons throughout the neostriatum are generated in a prominent ventrolateral-to-dorsomedial gradient so that ventrolateral cells originate mainly between E14 and E18, dorsomedial cells between E18 and E21–22 (fewer than 10% originate between P0 and P4). Medium-sized neurons also show two other gradients. First, there is a superficial-to-deep gradient in the anterior part of the caudoputamen, while more posterior levels have a deep-to-superficial gradient. Second, anterior parts have a caudal-to-rostral gradient while posterior parts have a gradient in the opposite direction. This shift in neurogenetic gradients along both superficial-deep and rostrocaudal directions is developmental evidence that an anterior ‘caudate’ can be separated from a posterior ‘putamen’ in the rat. Finally, neurogenetic gradients in the medium-sized caudoputamen neurons can be linked to the patterns of their anatomical interconnections with the substantia nigra.

3H-thymidine-radiographic studies of neurogenesis in the rat olfactory bulb.

Neurogenesis in the rat olfactory bulb was examined with 3H-thymidine-radiography. For the animals in the prenatal groups, the initial 3H-thymidine exposures were separated by 24 h; they were the offspring of pregnant females given two injections on consecutive embryonic (E) days (E12-E13, E13-E14, . . . E21-E22). For the animals in the postnatal (P) groups, the initial 3H-thymidine injections were separated by 48 h, each group receiving either four (PO-P3, P2-P4, . . . P6-P9) or two (P8-P9, P10-P11, . . . P20-P21) consecutive daily injections. On P60, the percentage of labeled cells and the proportion of cells added during either 24 h or 48 h periods were quantified at several anatomical levels for each neuronal population in the main olfactory bulb (mitral cells, tufted cells, granule cells, interneurons in the external plexiform layer, periglomerular granule cells) and accessory olfactory bulb (output neurons, granule cells, periglomerular granule cells). The total time span of neurogenesis extends from E12 to beyond P20. Output neurons are prenatally generated over 5-9 day periods (with most neurogenesis occurring over 2-4 days) in a strict sequential order beginning with the accessory bulb output neurons (E13-E14) and ending with the interstitial tufted cells lying between the glomeruli in the main bulb (E20-E22). These data are correlated with the main and accessory bulb projection fields in the amygdala and with the chronology of amygdala neurogenesis. With the exception of the granule cells in the accessory bulb (88% generated between E15-E22), the rest of the interneuronal populations are generated postnatally and nearly simultaneously. While most neurons (75-80%) originate during the first three weeks of life, all interneuronal populations, including accessory bulb granule cells, show some neurogenesis beyond P20. Injections of 3H-thymidine in juvenile and adult rats indicates neurogenesis up to P60 in the accessory bulb and up to P180 in the main bulb, especially in the main bulb granule cell population. There is circumstantial evidence for turnover of main bulb granule cells during adult life.

Quantitative 3H-thymidine radiographic analyses of neurogenesis in the rat amygdala.

AbstractNeurogenesis in the rat amygdala was examined with 3H‐thymidine radiography. The animals were the offspring of pregnant females given two injections of 3H‐thymidine on consecutive days in an overlapping series: Embryonic day (E) 12 + E13, + E14. E21 + E22. On 60 days of age, the percentage of labelled cells and the proportion of cells added during each day of formation were determined at several anatomical levels within the following components of the amygdala: anterior amygdaloid area, bed nuclei of the lateral and accessory olfactory tracts, central, medial, anterior cortical, posterolateral cortical, posteromedial cortical, basomedial, basolateral, and lateral nuclei, the amygdalo‐hippocampal area, and the intercalated masses.All large and many small neurons originate in most nuclei between E13 and E17, those in the intercalated masses between E15 and E19, those in the amygdalo‐hippocampal area between E16 and E19. The anterior amygdaloid area, intercalated masses, central, medial, posterolateral cortical, posteromedial cortical, basomedial, basolateral, and lateral nuclei have strong rostral‐to‐caudal intranuclear gradients. There are five additional intranuclear gradients: 1) medial to lateral in the central nucleus, anterior amygdaloid area, and anterior intercalated masses; 2) lateral to medial in the bed nucleus of the lateral olfactory tract and basolateral nucleus; 3) superficial to deep in the amygdalo‐hippocampal area, posterolateral, and posteromedial cortical nuclei; 4) ventral to dorsal in the medial nucleus; and 5) dorsal to ventral between the small and large‐celled parts of the lateral nucleus. Only the bed nucleus of the accessory olfactory tract and the anterior cortical nucleus do not have intranuclear gradients. Between 10 and 15% of the total cell population in most nuclei are very small neurons and/or glia which originate simultaneously between E18 and E20. This population is absent in the ventral part of the medial, anterior cortical, and anterior basomedial nuclei; these contiguous areas may form a distinct subunit in the amygdala.In contrast to the pronounced intranuclear gradient, internuclear gradients are weak. Neurogenesis in the central nucleus and corticomedial and basolateral complexes appears to take place both concurrently and independently. There are groups of early‐originating neurons in the central, medial, and basolateral nuclei located near the periphery of the amygdala. Each of these groups is surrounded by younger neurons farther within the interior. The youngest calls are in the centrally placed intercalated masses. These settling patterns suggest that cells in the amygdala arise simultaneously from more than one neuroepithelial source during morphogenesis.The chronology of neurogenesis in the amygdala can be related to some of its anatomical connections. Rostral‐to‐caudal gradients in the corticomedial complex may be timed to coincide with early vs. late arrival of olfactory fibers. The subdivisions bases on intranuclear gradients in the central, medial, and basolateral nuclei match the subdivisions based on patterns of anatomical connections.

Development of the hippocampal region in the rat I. Neurogenesis examined with 3H‐thymidine autoradiography

AbstractNeurogenesis in the rat hippocampal region was examined with 3H‐thymidine autoradiography. The rats in the prenatal groups were the offspring of pregnant females given two injections of 3H‐thymidine on consecutive days in an overlapping series: embryonic (E) day E13+E14, E14+E15,…, E21+E22. The rats in the postnatal (P) groups were injected in two nonoverlapping series: first, the day of birth (PO) and P1, P2+P3,…, P18+P19; second, P0–P3, P4–P7,…, P16–P19. On 60 days of age, the percentage of labelled cells and the proportion of cells added during each day of formation were determined at several anatomical levels within each structure of the hippocampal region (entorhinal cortex, parasubiculum, presubiculum, subiculum, Ammon's horn, and the dentate gyrus) and the hippocampal rudiment (tenia tecta, indusium griseum). The neurons in each structure arise in overlapping, but still significantly different, waves: the hippocampal rudiment between E16–E17; the entorhinal cortex between E15–E17; the para‐ and presubiculum between E16–E19; the subiculum between E16–E18; large cells in strata oriens, radiatum, lacunosum‐moleculare of Ammon's horn between E15–E17; Ammon's horn pyramidal cells between E17–E19; large cells in the dentate hilus and molecular layer between E15–E19. Dentate granule cells begin to originate on E17, and 10% of the population forms after P18.There are three characteristic gradients of formation within structures. First, deep cells are generated before superficial cells. Second, cells closer to the rhinal fissure are formed before those lying farther away (“rhinal to dentate” gradient). Third, later forming cells are flanked by earlier forming superficial and deep cells (“sandwich gradient”) in the entorhinal cortex (layer III cells originate after layers II and IV), Ammon's horn (pyramidal cells originate after large cells in strata oriens, radiatum, and lacunosum‐moleculare), and the dentate gyrus (granule cells originate after large cells in the hilus and molecular layer). There is a “rhinal to dentate” gradient between structures. The entorhinal cortex starts first, next is the subiculum, then field CA3 of Ammon's horn, and finally, the dentate gyrus. Two structures are exceptions to this gradient. The para‐ and presubiculum form significantly later than the subiculum, and CA1 forms significantly later than adjacent CA3 cells; this late neurogenesis may be related to prominent thalamic input to both structures.Neurogenetic gradients between the cells providing laminated afferent input to the Ammonic pyramidal and dentate granule cells correlate with their order of termination: afferents from progressively later‐originating cells terminate progressively closer to the cell body. Topographic hippocampal projections along the dorsoventral axis correlate with formation patterns in target structures: dorsal hippocampal fibers project to zones occupied by earlier‐forming cells in the lateral septal nucleus and pars posterior of the mammillary body; ventral hippocampal fibers project to zones occupied by later‐forming cells in these structures.