Postnatal Interval Research Articles

Neonatal gut and respiratory microbiota: coordinated development through time and space

BackgroundPostnatal development of early life microbiota influences immunity, metabolism, neurodevelopment, and infant health. Microbiome development occurs at multiple body sites, with distinct community compositions and functions. Associations between microbiota at multiple sites represent an unexplored influence on the infant microbiome. Here, we examined co-occurrence patterns of gut and respiratory microbiota in pre- and full-term infants over the first year of life, a period critical to neonatal development.ResultsGut and respiratory microbiota collected as longitudinal rectal, throat, and nasal samples from 38 pre-term and 44 full-term infants were first clustered into community state types (CSTs) on the basis of their compositional profiles. Multiple methods were used to relate the occurrence of CSTs to temporal microbiota development and measures of infant maturity, including gestational age (GA) at birth, week of life (WOL), and post-menstrual age (PMA). Manifestation of CSTs followed one of three patterns with respect to infant maturity: (1) chronological, with CST occurrence frequency solely a function of post-natal age (WOL), (2) idiosyncratic to maturity at birth, with the interval of CST occurrence dependent on infant post-natal age but the frequency of occurrence dependent on GA at birth, and (3) convergent, in which CSTs appear first in infants of greater maturity at birth, with occurrence frequency in pre-terms converging after a post-natal interval proportional to pre-maturity. The composition of CSTs was highly dissimilar between different body sites, but the CST of any one body site was highly predictive of the CSTs at other body sites. There were significant associations between the abundance of individual taxa at each body site and the CSTs of the other body sites, which persisted after stringent control for the non-linear effects of infant maturity. Canonical correlations exist between the microbiota composition at each pair of body sites, with the strongest correlations between proximal locations.ConclusionThese findings suggest that early microbiota is shaped by neonatal innate and adaptive developmental responses. Temporal progression of CST occurrence is influenced by infant maturity at birth and post-natal age. Significant associations of microbiota across body sites reveal distal connections and coordinated development of the infant microbial ecosystem.

Reducing endogenous estrogen during prepuberal life does not affect boar libido or sperm fertilizing potential

Increasing sperm production per breeding male has economic significance with increasing use of artificial insemination. Manipulations to increase sperm production in livestock will only be useful if libido and sperm fertilizing capacity are not adversely affected. Reducing endogenous estrogens in the postnatal interval increases the number of Sertoli cells and hence testicular sperm production capacity. These experiments were designed to evaluate the effects of reducing endogenous estrogens on libido and sperm fertilizing capacity. Boars were treated with an aromatase inhibitor, letrozole, to reduce testicular estrogen production between 1 and 6 weeks of age or between 11 and 16 weeks of age, and the littermates to these boars were treated with the canola oil vehicle. Letrozole treatment did not affect time to first mount at 22 weeks of age, regardless of whether the treatment occurred from 1 to 6 weeks of age (118 seconds vs. 233 seconds, SEM = 161 for letrozole-treated and vehicle-treated boars, respectively) or from 11 to 16 weeks of age (107 seconds vs. 67 seconds, SEM = 63 for letrozole-treated and vehicle-treated boars, respectively). Similarly, sperm fertilizing ability and in vivo fertility were equivalent in letrozole-treated boars and their vehicle-treated littermates. Surprisingly, the increase in Sertoli cell numbers observed in the letrozole-treated boars at 20 weeks of age (5.8 vs. 4.3 billion, SEM = 0.5; P < 0.05) was not maintained to 40 weeks of age in their letrozole-treated littermates. Reducing endogenous estrogen production neonatally or prepuberally had no detectable adverse effect on libido or sperm fertilizing capacity.

The relationship between neonatal developmental status and post-natal nutritional status in Hungarian children

Background: Neonatal development may have an influence on post-natal nutritional status. Age at adiposity rebound is critical for later development of nutritional status.Aim: The objective was to analyse the relationship between neonatal development and post-natal changes in nutritional status.Subjects and methods: Subjects were studied in a longitudinal national survey (1980–2001) from birth (n = 6219) to 18.0 years (n = 1448). Subjects were divided into small (SGA), appropriate (AGA) and large for gestational age (LGA) sub-groups. Nutritional status was assessed by BMI. The Reed--Asefa model was fitted to the subject’s serial data of BMI.Results: The body parameters of the neonatal developmental sub-groups differed significantly in all studied neonatal body dimensions: the higher the intra-uterine growth rate (the slowest growth rate was assumed in the SGA, the fastest in the LGA children), the heavier the body weight, the longer the length and the bigger the BMI values found. The nutritional status of the neonatal developmental sub-groups differed significantly during the studied post-natal interval: the higher the pre-natal growth rate, the better nutritional status (the larger BMI) was found after birth.Conclusions: Neonatal development influenced strongly the post-natal nutritional status of children. The results indicate that not only age at adiposity rebound but also neonatal developmental status can be used as an indicator of later obesity.

Early Infant Diet and the Omega 3 Fatty Acid DHA: Effects on Resting Cardiovascular Activity and Behavioral Development During the First Half-Year of Life

This investigation evaluated variations in resting heart rate (HR) measures during the first half year of life in healthy, full-term infants who were either breast-fed (BF), or fed formula with (milk-based: MF; soy-based: SF) or without (soy-based: SF−) commercially supplemented DHA (decosahexaenoic acid). In infants fed the DHA-deficient diet, higher HR and lower values for heart rate variability measures were observed, indicating decreased parasympathetic tone in this group. These effects, appearing at 4 months and continuing for the remainder of the study period, are consistent with suggestions that the 3–5-month postnatal interval may be an important period in the development of cardiovascular regulation. The absence of these effects in SF infants receiving the DHA-supplemented formula suggests that neither soy protein nor the associated phytochemicals in soy formula contribute to these effects to any appreciable extent. In general, the results do not indicate differences in any of the study variables attributable to soy formula per se.

Pias3-Dependent SUMOylation Directs Rod Photoreceptor Development

Specification of retinal rod photoreceptors is determined by several different transcription factors that activate expression of rod-specific genes and repress expression of cone photoreceptor-specific genes. The mechanism by which this dual regulation occurs is unclear. We have found that Pias3, a transcriptional coregulator and E3 SUMO ligase that is selectively expressed in developing photoreceptors, promotes the differentiation of rod photoreceptors while preventing rods from adopting cone photoreceptor-like characteristics. Pias3 binds the photoreceptor-specific transcription factors Crx and Nr2e3 and is specifically targeted to the promoters of photoreceptor-specific genes. Pias3 SUMOylates Nr2e3, converting it into a potent repressor of cone-specific gene expression. Rod- and cone-specific promoters are bound by hyperSUMOylated proteins in rod photoreceptors, and blocking SUMOylation in photoreceptors results in cells with morphological and molecular features of cones and an absence of rod-specific markers. Our data thus identify Pias3-mediated SUMOylation of photoreceptor-specific transcription factors as a key mechanism of rod specification.

Reducing endogenous estrogens during the neonatal and juvenile periods affects reproductive tract development and sperm production in postpuberal boars

Increased Sertoli cell proliferation during the neonatal period, transient negative effects on epididymal sperm maturation, larger postpuberal testis size reflective of increased Sertoli cell numbers, and increased testicular sperm production characterized boars subjected to continuous inhibition of endogenous estrogen production. The objective in the present experiment was to extend these previous observations to evaluate the effects of a shorter period of reduced estrogen production during the neonatal and juvenile periods on Sertoli cell proliferation, postpuberal testis size, sperm production and epididymal function. Experiments were designed to detect cumulative effects on accessory sex glands as well. Four pairs of littermate boars were assigned to the experiment with one member of each pair randomly selected to receive weekly oral treatment with the aromatase inhibitor, Letrozole, beginning at 1 week of age; the littermates received weekly oral treatment with the corn oil vehicle. Treatment stopped at 12 weeks of age and effects were examined at 10 months. Treated animals had approximately 25% larger testes ( P < 0.05) correlated with increased Sertoli cell numbers ( P < 0.05) and larger epididymides. Sperm quality was approximately equivalent in treated and control littermates. Accessory sex glands tended to be smaller in the treated boars. Sertoli cell proliferation during the neonatal and juvenile interval appears to be influenced by endogenous estrogen levels in the boar. A relatively short postnatal interval of Letrozole treatment effectively increased postpuberal testis size. Increased sperm production capacity in response to decreased endogenous estrogens has intriguing possibilities for animal agricultural production.

Chloride channelopathy in myotonic dystrophy resulting from loss of posttranscriptional regulation for CLCN1

Transmembrane chloride ion conductance in skeletal muscle increases during early postnatal development. A transgenic mouse model of myotonic dystrophy type 1 (DM1) displays decreased sarcolemmal chloride conductance. Both effects result from modulation of chloride channel 1 (CLCN1) expression, but the respective contributions of transcriptional vs. posttranscriptional regulation are unknown. Here we show that alternative splicing of CLCN1 undergoes a physiological splicing transition during the first 3 wk of postnatal life in mice. During this interval, there is a switch to production of CLCN1 splice products having an intact reading frame, an upregulation of CLCN1 mRNA encoding full-length channel protein, and an increase of CLCN1 function, as determined by patch-clamp analysis of single muscle fibers. In a transgenic mouse model of DM1, however, the splicing transition does not occur, CLCN1 channel function remains low throughout the postnatal interval, and muscle fibers display myotonic discharges. Thus alternative splicing is a posttranscriptional mechanism regulating chloride conductance during muscle development, and the chloride channelopathy in a transgenic mouse model of DM1 results from a failure to execute a splicing transition for CLCN1.

Failure of MBNL1-dependent post-natal splicing transitions in myotonic dystrophy

In myotonic dystrophy (DM), expression of RNA containing expanded CUG or CCUG repeats leads to misregulated alternative splicing of pre-mRNA. The repeat-bearing transcripts accumulate in nuclear foci, together with proteins in the muscleblind family, MBNL1 and MBNL2. In transgenic mice that express expanded CUG repeats, we show that the splicing defect selectively targets a group of exons that share a common temporal pattern of developmental regulation. These exons undergo a synchronized splicing switch between post-natal day 2 and 20 in wild-type mice. During this post-natal interval, MBNL1 protein translocates from a predominantly cytoplasmic to nuclear distribution. In the absence of MBNL1, these physiological splicing transitions do not occur. The splicing defect induced by expanded CUG repeats in mature muscle fibers is closely reproduced by deficiency of MBNL1 but not by deficiency of MBNL2. A parallel situation exists in human DM type 1 and type 2. MBNL1 is depleted from the muscle nucleoplasm because of sequestration in nuclear foci, and the associated splicing defects are remarkably similar to those observed in MBNL1 knockout mice. These results indicate that MBNL1 participates in the post-natal remodeling of skeletal muscle by controlling a key set of developmentally regulated splicing switches. Sequestration of MBNL1, and failure to maintain these splicing transitions, has a pivotal role in the pathogenesis of muscle disease in DM.

Motor, cytoarchitectural and biochemical assessment of pharmacological neuroprotection against CNS damage induced by neonatal exposure to ionizing radiation

Exposure of neonatal rats to a 5 Gy single dose of X-irradiation induces permanent abnormalities in cerebellar cortex cytoarchitecture and neurochemistry and motor function. This rodent model constitutes an useful tool to evaluate morphological, neurochemical and motor changes induced by ionizing radiation and the possible restorative effects of potential or clearly established neuroprotective drugs. After selection and administration of a neuroprotective agent to neonatally irradiated rats, quantitative evaluations of motor behavior (gait), cerebellar cortex cytoarchitecture and cerebellar monoamine levels are performed. Data are compared to those of both saline-injected, X-irradiated, and saline-injected, sham-irradiated controls. Evaluation of data from the different experimental groups is performed at postnatal days 30 and 90. After this postnatal interval, radiation-induced damage of cerebellar function in nonprotected rodents is considered to be permanent. The longitudinal evaluation of various parameters in the different experimental groups through a multidisciplinary approach, allows determination of the variables that are more sensitive to X-irradiation-induced damage and/or neuroprotective agent-induced restoration. Given the well-known correspondence in cerebellar developmental stages between rodents and humans, this model and related studies bring health-related implications, considering the accidental or therapeutic exposure of developing human beings to ionizing radiation.

Sequence of Neuron Origin and Neocortical Laminar Fate: Relation to Cell Cycle of Origin in the Developing Murine Cerebral Wall

Neurons destined for each region of the neocortex are known to arise approximately in an "inside-to-outside" sequence from a pseudostratified ventricular epithelium (PVE). This sequence is initiated rostrolaterally and propagates caudomedially. Moreover, independently of location in the PVE, the neuronogenetic sequence in mouse is divisible into 11 cell cycles that occur over a 6 d period. Here we use a novel "birth hour" method that identifies small cohorts of neurons born during a single 2 hr period, i.e., 10-20% of a single cell cycle, which corresponds to approximately 1.5% of the 6 d neuronogenetic period. This method shows that neurons arising with the same cycle of the 11 cycle sequence in mouse have common laminar fates even if they arise from widely separated positions on the PVE (neurons of fields 1 and 40) and therefore arise at different embryonic times. Even at this high level of temporal resolution, simultaneously arising cells occupy more than one cortical layer, and there is substantial overlap in the distributions of cells arising with successive cycles. We demonstrate additionally that the laminar representation of cells arising with a given cycle is little if at all modified over the early postnatal interval of histogenetic cell death. We infer from these findings that cell cycle is a neuronogenetic counting mechanism and that this counting mechanism is integral to subsequent processes that determine cortical laminar fate.

Clinical effects of porcine reproductive and respiratory syndrome virus on pigs during the early postnatal interval

SUMMARY Objective To determine the effect of congenital and early postnatal infection of pigs with porcine reproductive and respiratory syndrome virus (PRRSV) on postnatal survival and growth. Animals 20 pregnant gilts and their pigs and fetuses. Procedure 16 pregnant gilts (principals) comprising 4 groups (4 gilts/group) were exposed oronasally to 4 strains of PRRSV (a vaccine strain, and 3 field strains) at or about day 90 of gestation. Four pregnant gilts (controls) were kept under similar conditions, except for exposure to PRRSV. Samples collected from pigs before ingestion of colostrum and samples and specimens collected from pigs at selected times thereafter were tested for PRRSV and homologous antibody. Pigs were observed for clinical signs and were weighed at birth and at weekly intervals until they were euthanatized and necropsied at about 3 weeks of age. Results At least some members of all litters of principal gilts were infected congenitally. Most noninfected, liveborn littermates became infected within the first week of life. Infection of pigs with field strains did, and infection of pigs with the vaccine strain did not, adversely affect postnatal survival and growth rate. All infected pigs had generalized lymph node enlargement. Conclusion Exposure of pregnant gilts to either attenuated (vaccine) or virulent (field) strains of PRRSV can result in congenital infection. Vaccine as well as field strains can be transmitted postnatally from infected to noninfected littermates. Pigs infected with field strains have a poorer rate of survival and growth than do noninfected pigs. Clinical Relevance Because attenuated (vaccine) PRRSV can cause congenital infection and be transmitted postnatally from congenitally infected to immune-naive pigs, the use of attenuated virus during gestation is, at best, questionable. (Am J Vet Res 1998;59:52–55)

Can the mechanisms of aluminum neurotoxicity be integrated into a unified scheme?

Regardless of the host, the route of administration, or the speciation, aluminum is a potent neurotoxicant. In the young adult or developmentally mature host, the neuronal response to Al exposure can be dichotomized on morphological grounds. In one, intraneuronal neurofilamentous aggregates are formed, whereas in the other, significant neurochemical and neurophysiological perturbations are induced without neurofilamentous aggregate formation. Evidence is presented that the induction of neurofilamentous aggregates is a consequence of alterations in the posttranslational processing of neurofilament (NF), particularly with regard to phosphorylation state. Although Al has been reported to impact on gene expression, this does not appear to be critical to the induction of cytoskeletal pathology. In hosts responding to Al exposure without the induction of cytoskeletal pathology, impairments in glucose utilization, agonist-stimulated inositol phosphate accumulation, free radical-mediated cytotoxicity, lipid peroxidation, reduced cholinergic function, and altered protein phosphorylation have been described. The extent to which these neurochemical modifications correlate with the induction of a characteristic neurobehavioral state is unknown. In addition to these paradigms, Al is toxic in the immediate postnatal interval. Whether unique mechanisms of toxicity are involved during development remains to be determined. In this article, the mechanisms of Al neurotoxicity are reviewed and recommendations are put forth with regard to future research. Primary among these is the determination of the molecular site of Al toxicity, and whether this is based on Al substitution for divalent metals in a number of biological processes. Encompassed within this is the need to further understand the genesis of host- and developmental-specific responses.

Transcriptional and post-transcriptional mechanisms regulating neurofilament and tubulin gene expression during normal development of the rat brain

The transcription of the beta II-, beta IV- and alpha 1-tubulin genes as well as that of the three neurofilament genes, NF-L, NF-M and NF-H, was examined during the course of postnatal brain development. Changes in the transcriptional activity of these genes were studied using run-off transcription assays with nuclei isolated from the rat cerebral cortex at postnatal days P2, P5, P10 and adult stages. Northern blotting of total RNA isolated from the cerebral cortex was used to compare changes in steady-state mRNA levels with transcriptional changes that occurred in the cerebral cortex during the postnatal interval. Nuclear run-off assays showed that beta II- and alpha 1-tubulin gene transcription rates were maximal from P2-P5 and declined at later times. Changes in the steady-state mRNA levels for these two genes followed the same general pattern as transcription, but in the case of beta II-tubulin mRNA, were more dramatic. beta IV-tubulin gene transcription dropped between P2 and P5 and then increased progressively to the adult stage, coordinate with an increase in beta IV-tubulin steady-state mRNA levels. NF-L and NF-H genes showed similar patterns of transcriptional change during the postnatal interval, with maximal rates of transcription at P5 followed by a decline at later times. The steady-state levels of NF-L and NF-H mRNAs changed in a manner opposite to that of transcription and increased progressively during the postnatal interval. This suggests that mRNA stabilization is the main factor regulating the steady-state levels of NF-L and NF-H mRNAs in postnatal brain. For the NF-M gene, the developmental transcription pattern was also dissociated from steady-state mRNA level changes, but differed from the transcription patterns of the NF-L and NF-H genes. This suggests the importance of post-transcriptional mechanisms in regulating NF-M mRNA levels in brain and also indicates that some differences exist in the regulatory mechanisms which control NF-M compared to NF-L and NF-H mRNA levels.

Coordinate regulation of tubulin and microtubule associated protein genes during development of hamster brain

The present study documents the patterns of mRNA expression for 5 different tubulin genes and 4 of the structural microtubule-associated protein (MAP) genes during normal development of hamster forebrain. Northern blotting in conjunction with densitometric analysis was used to study changes in the levels of the mRNAs for α I-tubulin, classes β I-, β II-, β III- and β IV-tubulin, as well as the mRNAs for tau, MAP1A, MAP1B and MAP2, using total RNA isolated from hamster forebrain at various embryonic (E) and postnatal (P) stages. Densitometric analyses of the autoradiograms from the Northern blots revealed that each of the tubulin genes exhibited distinct developmental patterns of expression, several of which appeared to be temporally correlated with the expression of specific MAP mRNAs. The β I-, β II- and β III-tubulin mRNAs increased rapidly between late embryonic stages to birth, reached peak levels early in the first postnatal week, and declined thereafter. α I-Tubulin mRNA was easily detected during embryonic stages, rose to peak levels at P7–P9 and then gradually declined. A similar pattern was seen for tau mRNA. After the first postnatal week, the size of the tau mRNA also shifted to a slightly larger size, presumably due to differential splicing. β IV-Tubulin mRNA levels did not become significant until very late in postnatal development (3–4 weeks). MAP2 mRNA expression was unusual in that peak levels were reached at two different stages of development - an initial peak occurred in the first postnatal week, followed by a decline, and then a second rise occurred during the third and fourth postnatal weeks. The expression of the β IV-tubulin mRNA coincided temporally with the second peak in MAP2 mRNA expression. MAP1B mRNA abruptly reached high levels at birth, remained abundant during the first two postnatal weeks, and then decreased. In contrast, MAP1A mRNA levels were low in the initial postnatal interval and increased only at very late developmental stages. The findings of a temporal correspondence in expression of high levels of tau and MAP1B with β I-, β II-, β III- and αI-tubulin mRNAs suggest that this profile of gene expression is one that endows greater plasticity to the neuronal cytoskeleton. Conversely, the transition to increased expression of β IV-tubulin, MAP1A, and a larger tau mRNA species defines a portion of the molecular pattern that underpins the increased stability of neuronal form during maturation.

Developmental patterns of intermediate filament gene expression in the normal hamster brain

We have examined the patterns of expression of the major intermediate filament (IF) protein mRNAs during development of the hamster brain. Quantitative northern blotting was used to examine changes in the levels of mRNAs for the low, middle and high molecular weight neurofilament proteins (NF-L, NF-M, NF-H) as well as peripherin, vimentin and glial fibrillary acidic protein (GFAP). Total RNA was isolated from hamster brains at embryonic (E) days 12 and 14 and postnatal (P) days 1, 3, 5, 7, 9, 11, 13, 15, 20, 28 and 60–90 (adult), and probed with specific IF cDNAs. Northern blotting revealed that NF-L and NF-M mRNAs were present at very low levels in embryonic brain and that significant expression of these genes only occurred postnatally when the levels increased dramatically until P28 and then declined again in the adult. Increases in NF-H mRNA levels were somewhat delayed relative to those of NF-L and NF-M. NF-H mRNA was not seen at embryonic stages and was expressed at very low levels prior to P9; after that time the levels increased rapidly until P28 and then declined in the adult. Two of the type III IF genes, peripherin and vimentin, followed a pattern of expression opposite that of the NF genes. Both peripherin and vimentin mRNAs were present in embryonic brain and were expressed at higher levels during early postnatal stages than at later times. The magnitude and rate of reduction in vimentin gene expression in the postnatal interval was much greater than that of peripherin. GFAP mRNA levels were extremely low prior to P9 alter which a robust increase occurred, followed by a decline in the adult. We discuss the implication of the dramatic changes in IF isotype expression in brain to the pathways of both neuronal and glial development in vivo.

An ecological theory for the origin ofHomo

The genusHomoevolved its pronounced encephalization through postnatal extension of the high rate of brain growth that characterizes all primatesin utero.Linked to this extension was delayed development, which represented an enormous ecological sacrifice because it produced the longest postnatal interval of physical helplessness in the Mammalia and forced mothers to carry infants.Graphs relating brain growth to body growth indicate a pongid pattern of development for gracile australopithecines, implying that infants could cling to mothers whose forelimbs were occupied with climbing. Also present were several postcranial traits that would have made the adults more adept climbers than modern humans. Habitual use of these inherited traits is suggested by the fact that evolution failed to eliminate certain ones, such as short legs and long pedal phalanges, that impeded terrestrial locomotion. Moreover, the intensity of predation by large, swift, social carnivores must have compelled australopithecines to use trees as refuges, in the manner of chimpanzees and baboons; australopithecines probably also gathered some of their food in trees. Gracile australopithecines failed to expand their brain size, experiencing general evolutionary stasis for more than 1.5 m.y. I propose that this stability resulted from these animals' semiarboreal mode of life: First, their postcranial morphology remained compromised by selection pressures to maintain both terrestrial and arboreal adaptations. Second, by requiring that neonates be mature enough to cling to mothers, obligate arboreal activity precluded encephalization of the kind that characterizesHomo;this evolutionary constraint has previously been overlooked.In contrast to australopithecines, earlyHomoapproachedH. erectusin pelvic configuration and brain size. A new brain-body growth curve for earlyHomoindicates extension of the fetal pattern well into the postnatal interval, implying that neonates were highly immature so that adults had to be fully terrestrial.Homoevolved shortly after the onset of the modern ice age about 2.5 Ma. Fossil pollen and carbon isotopes in paleosols record a contraction of forests in Africa at this time. I propose that this represented a crisis that led to the evolution ofHomoby compelling some australopithecine populations to adopt a fully terrestrial existence. Although ecologically difficult, this behavioral restriction finally made possible encephalization through the evolution of delayed development. During the ecological crisis, a large brain evolved in at least one population of gracile australopithecines. An advanced tool industry and cunning behavior were of such great adaptive value for avoiding predators and expanding food resources on the ground that selection for encephalization soon overrode the problems imposed by helpless infants.The fossil record of antelopes and micromammals provides a test of the idea that environmental forcing opened the way for the evolution ofHomo:both of these groups experienced heavy extinction of forest-adapted species about 2.5–2.4 Ma and a rapid proliferation of species adapted to unforested habitats. The transformation of the hominid clade during Plio-Pleistocene time did not follow a simple pattern.Homomay have arisen either by anagenetic transformation of a “bottlenecked” species or by speciation, and it may not have evolved immediately with the onset of climatic change. Furthermore, just as a few forest-adapted antelope species survived the biotic crisis, a small-brained gracile taxon with arboreal adaptations may have persisted to the start of the Pleistocene. Robust australopithecines survived into the Pleistocene, perhaps because a broad vegetarian diet reduced their need to migrate frequently between home bases. With their extinction in mid-Pleistocene time, about the time that savannahs became widespread, onlyHomoremained.

Ontogenetic analysis of some surface markers on pig lymphocytes using fluorescence-activated cell sorter

Surface markers were demonstrated on pig lymphocytes using anti-T cell-IgG and anti-Helix pomatia (HP) IgG during prenatal and postnatal development. A fluorescence-activated cell sorter analysis of T-cell surface markers was accompanied by an image analysis to prove the association of T antigenic determinants with the plasma membrane only. We found development-dependent changes in both anti-T cell and HP surface markers in both primary and secondary lymphatic organs. The number of T-positive (T+) cells estimated by anti-T cell-IgG was very similar to the results obtained by spontaneous E-rosette forming tests. At all selected age intervals, changes in the number of T+ cells were not significant in the thymus, but a marked increase in T+ cells was found in both spleen and lymph nodes. The image analysis confirmed the expression of T cell markers on the cell surface. The distribution of T cell markers was uneven, i.e. various degree of fluorescence intensity on whole ring-pattern projection of the cell surface image was estimated. In second lymphatic organs especially, fluorescence intensity of cells, i.e. total number of T cell markers estimated by anti-T cell-IgG, increased with age. On fetal day 73, T cell markers were slightly expressed, but very high fluorescence intensity and heterogeneous distribution of T cell markers on lymphocytes were found on fetal day 107 and postnatal day 56. The results indicate the possibility of functional maturation of various T cell markers on T cell subsets, furthermore a different degree of expression of T cell markers on various T cell subsets can be suggested. The number of HP+ cells increased with age in both primary and secondary lymphatic organs. In the prenatal period, the expression of HP receptors was very weak in both primary and secondary organs in contrast to the marked increase in HP+ cells during the postnatal interval. Differences in fluorescence intensity of cells were found, representing the increase by 22% in thymus cells comparing to cells of secondary lymphatic organs. Heterogeneity of HP+ cell populations in thymus was shown by the Scatchard plot, indicating at least two subpopulations of HP+ cells with different avidity to HP. Cells with low HP avidity could include a subset with cytolytic activity.

Postnatal dendritic development in motoneurons: evaluation by a Monte Carlo technique.

A Monte Carlo method was used to quantitate the dendritic characteristics of horseradish peroxidase-injected lumbar motoneurons (MNs) from kittens 44 to 73 days old (during the postnatal interval when motor cortical input/output linkages mature) and compared with those from adults. All of the 6 MNs analyzed were within the same range of somal sizes (40-61 micron) and dendritic domain volumes. However, two-dimensional distributions of dendritic diameter versus distance from the soma revealed that in the youngest MNs' (44 and 51 days) most dendritic processes (82 and 84%) were less than 2 micron in diameter, and had no dendrites greater than 5 micron. The adult MNs had large dendrites (20 and 24% were greater than 5 mu), while the kittens of intermediate age (66 and 73 days) exhibited both adult-like (6 and 13% greater than 5 micron) and immature (48 and 73% less than 2 micron) characteristics. "Hit probabilities" calculated from the Monte Carlo data suggest that immature MNs' dendrites may present denser targets per unit volume for ingrowing growth cones than adult MNs, and may facilitate the probability of contact with appropriate afferent axons.

Development of beta-adrenergic receptors in normal and mutant mouse cerebellum.

[3H]-dihydroalprenolol was used to examine the development of beta-adrenoceptors in the cerebellum of weaver, reeler, staggerer, and jimpy neurologically mutant mice and their normal counterparts. In normal animals the greatest increase in [3H] binding occurred during the postnatal interval of 10-15 days, and maximum values were obtained at about 3 weeks. Binding was saturable with an apparent dissociation constant of 0.26 nM, and the affinity of [3]-dihydroalprenolol for its receptor did not change significantly during development. [3H]-dihydroalprenolol binding was significantly reduced in weaver, reeler, and staggerer (14-22% of control values) at 20 days, but not in the jimpy mutant. These results are discussed in relation to the ontogeny of beta-adrenoreceptors with the known noradrenergic innervation of the cerebellum, and to the paucity of both granule neurons and myelination which occurs in the neurological mutants.