Weeks Of Postnatal Development Research Articles

Molecular and functional characterization of GAD67-expressing, newborn granule cells in mouse dentate gyrus

Dentate gyrus granule cells (GCs) have been suggested to synthesize both GABA and glutamate immediately after birth and under pathological conditions in the adult. Expression of the GABA synthesizing enzyme GAD67 by GCs during the first few weeks of postnatal development may then allow for transient GABA synthesis and synaptic release from these cells. Here, using the GAD67-EGFP transgenic strain G42, we explored the phenotype of GAD67-expressing GCs in the mouse dentate gyrus. We report a transient, GAD67-driven EGFP expression in differentiating GCs throughout ontogenesis. EGFP expression correlates with the expression of GAD and molecular markers of GABA release and uptake in 2–4 weeks post-mitotic GCs. These rather immature cells are able to fire action potentials (APs) and are synaptically integrated in the hippocampal network. Yet they show physiological properties that differentiate them from mature GCs. Finally, GAD67-expressing GCs express a specific complement of GABAA receptor subunits as well as distinctive features of synaptic and tonic GABA signaling. Our results reveal that GAD67 expression in dentate gyrus GCs is a transient marker of late differentiation that persists throughout life and the G42 strain may be used to visualize newborn GCs at a specific, well-defined differentiation stage.

Molecular cloning and localization of a CEACAM2 isoform, CEACAM2‐L, expressed in spermatids in mouse testis

Carcinoembryonic antigen (CEA) family, a subgroup of the immunoglobulin (Ig) superfamily, is divided into two sub-families: the CEA-related cell adhesion molecules (CEACAM) and the pregnancy-specific glycoproteins. The isoform CEACAM2 is expressed in mouse testis; in this study, we identified a novel isoform of Ceacam2, Ceacam2-Long (Ceacam2-L). CEACAM2-L is different from CEACAM2 in that it has much longer cytoplasmic tail region. Ceacam2-L starts to appear faintly in mouse testis after 3 weeks of postnatal development, and its expression level increased after 5 weeks. Immunoblot analysis confirmed the expression of CEACAM2-L in the seminiferous epithelium of mouse testis. Immunohistochemical data showed that CEACAM2-L was not observed on spermatogonia, spermatocytes, round spermatids, or Sertoli cells, but was seen at the plasma membrane of elongating spermatids in contact with extended cytoplasmic processes of Sertoli cells. CEACAM2-L was not detected at the head region of elongating spermatids, where the apical ectoplasmic specialization is constructed. These data suggest that CEACAM2-L might be a novel adhesion molecule contributing to cell-to-cell adhesion between elongating spermatids and Sertoli cells within the seminiferous epithelium.

Pur‐alpha regulates RhoA developmental expression and downstream signaling

Pur-alpha is an essential protein for postnatal brain development which localizes specifically to dendrites where it plays a role in the translation of neuronal RNA. Mice lacking Pur-alpha display decreased neuronogenesis and impaired neuronal differentiation. Here we examined two Rho GTPases, Rac1 and RhoA, which play opposing roles in neurite outgrowth and are critical for dendritic maturation during mouse brain development in the presence and absence of Pur-alpha. Pur-alpha is developmentally regulated in the mouse brain with expression beginning shortly after birth and rapidly increasing to peak during the third week of postnatal development. RhoA levels analyzed by Western blotting rapidly fluctuated in the wild-type mouse brain, however, in the absence of Pur-alpha, a decrease in RhoA levels shortly after birth and a delay in the cycling of RhoA regulation was observed leading to reduced basal levels of RhoA after day 10 postnatal. Immunohistochemistry of brain tissues displayed reduced RhoA levels in the cortex and cerebellum and loss of perinuclear cytoplasmic labeling of RhoA within the cortex in the knockout mouse brain. While Rac1 levels remained relatively stable at all time points during development and were similar in both wild-type and Pur-alpha knockout mice, changes in subcellular localization of Rac1 were seen in the absence of Pur-alpha. These findings suggest that Pur-alpha can regulate RhoA at multiple levels including basal protein levels, subcellular compartmentalization, as well as turnover of active RhoA in order to promote dendritic maturation.

Effects of Caudal Autotomy on Postnatal Growth Rates of HatchlingPsammodromus algirus

Abstract Tail autotomy has clear advantages regarding predator escape, but it also has several associated costs (i.e., impaired locomotion, loss of social status, and reduced growth and reproductive output). We examined the costs of severe autotomy on growth rates of hatchlings of the lacertid lizard Psammodromus algirus during the first weeks of postnatal development. Hatchlings from two populations in central Spain were autotomized on the fifth day after hatching and kept in common garden conditions for 35 days when they were measured again. Hatchlings from both populations, independently of the autotomy treatment, did not differ in the mass gained during the experiment. However, there were differences in body growth between tailless and tailed hatchlings; tailless hatchlings grew at a slower rate than tailed ones, after controlling for the effects of body condition at the onset of the experiment and the resources assimilated. Moreover, independently of their population of origin, hatchlings that invest...

Postnatal subventricular zone of the neocortex contributes GFAP+ cells to the rostral migratory stream under the control of Sip1

The rostral migratory stream (RMS) is composed of neuroblasts migrating from the striatal SVZ to the olfactory bulb through a meshwork of GFAP- expressing astrocytes called the glial tube. So far, the origin of the glial tube astrocytes was attributed to differentiation of Type-B stem cells of the striatal SVZ. The true identity of these cells (Type-B stem cells versus immature/mature astrocytes) is also unclear. By analyzing a neocortex-specific conditional knockout of the transcriptional repressor Sip1 (Smad-interacting protein 1), we have now identified a novel pool of progenitors located within the dorsal SVZ (dSVZ) at early postnatal stages that differentiate into GFAP+ cells of the glial tube. We show that Sip1, expressed in postmitotic cortical neurons, controls the size of this dorsal progenitor pool possibly through cell-extrinsic mechanisms. Lack of Sip1 in the neocortex causes an expansion of this population leading to an increased production of GFAP+ astrocytes/Type-B stem cells in the glial tube, and a denser intercalation of these cells with Dcx+ neuroblasts of the RMS, the consequence of which is not yet clear. Neocortex-specific Sip1 deletion also led to an expansion of Dcx+ and Tbr2+ progenitor populations in the dSVZ. We show that the dSVZ progenitors (possibly remnants of embryonic radial glia) differentiate exclusively into BLBP+ cells which migrate into the RMS and mature into GFAP+ astrocytes/Type-B stem cells at around two weeks of postnatal development. In summary, our work shows that Sip1 controls the generation of GFAP+ cells of the RMS by regulating the size of a novel progenitor pool located in the postnatal dSVZ.

Local generation of glia is a major astrocyte source in postnatal cortex

Glial cells constitute nearly 50% of the cells in the human brain. Astrocytes, which make up the largest glial population, are crucial to the regulation of synaptic connectivity during postnatal development. Because defects in astrocyte generation are associated with severe neurological disorders such as brain tumours, it is important to understand how astrocytes are produced. Astrocytes reportedly arise from two sources: radial glia in the ventricular zone and progenitors in the subventricular zone, with the contribution from each region shifting with time. During the first three weeks of postnatal development, the glial cell population, which contains predominantly astrocytes, expands 6-8-fold in the rodent brain. Little is known about the mechanisms underlying this expansion. Here we show that a major source of glia in the postnatal cortex in mice is the local proliferation of differentiated astrocytes. Unlike glial progenitors in the subventricular zone, differentiated astrocytes undergo symmetric division, and their progeny integrate functionally into the existing glial network as mature astrocytes that form endfeet with blood vessels, couple electrically to neighbouring astrocytes, and take up glutamate after neuronal activity.

Regulation of connexin 36 expression during development

In the mammalian CNS, the expression of neuronal gap junction protein, connexin 36 (Cx36), increases during the first 2 weeks of postnatal development and then decreases during the following 2 weeks. Recently we showed that the developmental increase in Cx36 expression is augmented by chronic (2 weeks) activation of group II metabotropic glutamate receptors (mGluR), prevented by chronic receptor inactivation, and the receptor-dependent increase in Cx36 expression is regulated via transcriptional control of the Cx36 gene activity. We demonstrate here that acute (60min) activation of group II mGluRs in developing cortical neuronal cultures causes transient increase in Cx36 protein expression with decrease during the following 24h. However, there is no change in Cx36 mRNA expression. In addition, the data indicate that transient increase in Cx36 expression is due to new protein synthesis. The results suggest that, during development, acute activation of group II mGluRs causes up-regulation of Cx36 via post-transcriptional mechanisms. However, if the receptor activation is sustained, transcriptional activation of the Cx36 gene occurs.

Growth Defects and Impaired Cognitive–Behavioral Abilities in Mice with Knockout for Eif4h, a Gene Located in the Mouse Homolog of the Williams-Beuren Syndrome Critical Region

Protein synthesis is a tightly regulated, energy-consuming process. The control of mRNA translation into protein is fundamentally important for the fine-tuning of gene expression; additionally, precise translational control plays a critical role in many cellular processes, including development, cellular growth, proliferation, differentiation, synaptic plasticity, memory, and learning. Eukaryotic translation initiation factor 4h (Eif4h) encodes a protein involved in the process of protein synthesis, at the level of initiation phase. Its human homolog, WBSCR1, maps on 7q11.23, inside the 1.6 Mb region that is commonly deleted in patients affected by the Williams-Beuren syndrome, which is a complex neurodevelopmental disorder characterized by cardiovascular defects, cerebral dysplasias and a peculiar cognitive-behavioral profile. In this study, we generated knockout mice deficient in Eif4h. These mice displayed growth retardation with a significant reduction of body weight that began from the first week of postnatal development. Neuroanatomical profiling results generated by magnetic resonance imaging analysis revealed a smaller brain volume in null mice compared with controls as well as altered brain morphology, where anterior and posterior brain regions were differentially affected. The inactivation of Eif4h also led to a reduction in both the number and complexity of neurons. Behavioral studies revealed severe impairments of fear-related associative learning and memory formation. These alterations suggest that Eif4h might contribute to certain deficits associated with Williams-Beuren syndrome.

Development of the sigma-1 receptor in C-terminals of motoneurons and colocalization with the N,N′-dimethyltryptamine forming enzyme, indole-N-methyl transferase

The function of the sigma-1 receptor (S1R) has been linked to modulating the activities of ion channels and G-protein-coupled receptors (GPCR). In the CNS, the S1R is expressed ubiquitously but is enriched in mouse motoneurons (MN), where it is localized to subsurface cisternae of cholinergic postsynaptic densities, also known as C-terminals. We found that S1R is enriched in mouse spinal MN at late stages of embryonic development when it is first visualized in the endoplasmic reticulum. S1Rs appear to concentrate at C-terminals of mouse MN only on the second week of postnatal development. We found that indole-N-methyl transferase (INMT), an enzyme that converts tryptamine into the sigma-1 ligand dimethyltryptamine (DMT), is also localized to postsynaptic sites of C-terminals in close proximity to the S1R. This close association of INMT and S1Rs suggest that DMT is synthesized locally to effectively activate S1R in MN.

Ubiquitin Homeostasis Is Critical for Synaptic Development and Function

The ubiquitin-proteasome system (UPS) controls protein abundance and is essential for many aspects of neuronal function. In ataxia (ax(J)) mice, profound neurological and synaptic defects result from a loss-of-function mutation in the proteasome-associated deubiquitinating enzyme Usp14, which is required for recycling ubiquitin from proteasomal substrates. Here, we show that transgenic complementation of ax(J) mice with neuronally expressed ubiquitin prevents early postnatal lethality, restores muscle mass, and corrects developmental and functional deficits resulting from the loss of Usp14, demonstrating that ubiquitin deficiency is a major cause of the neurological defects observed in the ax(J) mice. We also show that proteasome components are normally induced during the first 2 weeks of postnatal development, which coincides with dramatic alterations in polyubiquitin chain formation. These data demonstrate a critical role for ubiquitin homeostasis in synaptic development and function, and show that ubiquitin deficiency may contribute to diseases characterized by synaptic dysfunction.

Delayed synapse elimination in mouse levator palpebrae superioris muscle

At birth, synaptic sites in developing rodent muscles are innervated by numerous motor axons. During subsequent weeks, this multiple innervation disappears as one terminal strengthens, and all the others are eliminated. Experimental perturbations that alter neuromuscular activity affect the rate of synaptic refinement, with more activity accelerating the time to single innervation and neuromuscular blockade retarding it. However, it remains unclear whether patterns of muscle use (driven by endogenous neuronal activity) contribute to the rate of synapse elimination. For this reason we examined the timing of supernumerary nerve terminal elimination at synapses in extraocular muscles (EOMs), a specialized set of muscles controlling eye movements. On the basis of their exceptionally high patterns of activity, we hypothesized that synaptic refinement would be greatly accelerated at these synapses. We found, however, that rates of synaptic refinement were only modestly accelerated in rectus and oblique EOMs compared with synapses in somite-derived skeletal muscle. In contrast to these results, we observed a dramatic delay in the elimination of supernumerary nerve terminals from synapses in the levator palpebrae superioris (LPS) muscle, a specialized EOM that initiates and maintains eyelid elevation. In mice, natural eye opening occurs at the end of the second postnatal week of development. Thus, although synapse elimination is occurring in most EOMs and somite-derived skeletal muscles, it appears to be dramatically delayed in a set of specialized eyelid muscles that remain immobile during early postnatal development.

Delayed synapse elimination in mouse levator palpebrae superioris muscle

AbstractAt birth, synaptic sites in developing rodent muscles are innervated by numerous motor axons. During subsequent weeks, this multiple innervation disappears as one terminal strengthens, and all the others are eliminated. Experimental perturbations that alter neuromuscular activity affect the rate of synaptic refinement, with more activity accelerating the time to single innervation and neuromuscular blockade retarding it. However, it remains unclear whether patterns of muscle use (driven by endogenous neuronal activity) contribute to the rate of synapse elimination. For this reason we examined the timing of supernumerary nerve terminal elimination at synapses in extraocular muscles (EOMs), a specialized set of muscles controlling eye movements. On the basis of their exceptionally high patterns of activity, we hypothesized that synaptic refinement would be greatly accelerated at these synapses. We found, however, that rates of synaptic refinement were only modestly accelerated in rectus and oblique EOMs compared with synapses in somite‐derived skeletal muscle. In contrast to these results, we observed a dramatic delay in the elimination of supernumerary nerve terminals from synapses in the levator palpebrae superioris (LPS) muscle, a specialized EOM that initiates and maintains eyelid elevation. In mice, natural eye opening occurs at the end of the second postnatal week of development. Thus, although synapse elimination is occurring in most EOMs and somite‐derived skeletal muscles, it appears to be dramatically delayed in a set of specialized eyelid muscles that remain immobile during early postnatal development. J. Comp. Neurol. 519:2907‐2921, 2011. � 2011 Wiley‐Liss, Inc.

Time-of-day dependence of neurological deficits induced by sodium nitroprusside in young mice

Sodium nitroprusside (SNP) is widely used in pharmacological studies as a potent vasodilator or a nitric oxide donor. SNP-induced ataxic effects were assessed in mice by the Joulou-Couvoisier test. Swiss albino mice of both genders, 2-8 weeks of age, were acclimated at least for 2 weeks to 12 h light (rest span)/12 h dark (activity span). In 2 and 4 week old mice, maxima of ataxia were found following intraperitoneal administration of a dose ranging from 3 to 3.6 mg.kg-1 SNP at ≈ 1 and 13 HALO (Hours After Light Onset). The sublethal toxicity was statistically dosing-time dependent (χ2 test: P < 0.005). No rhythm was validated in neurotoxicity by cosinor analyses. At the 8th week of post-natal development (PND), SNP-induced ataxia was greatest at ≈ 1 HALO (69% in males vs. 49% in females) and lowest at ≈ 13 HALO (21% in males vs. 11% in females) (χ2 test: P < 0.00001). Cosinor analysis also revealed no statistically significant rhythm in mice injected with 3 or 3.3 mg.kg-1. However, a significant circadian (τ = 24 h) rhythm was detected by adjusted cosinor in 3.6 mg.kg-1-treated mice (P < 0.004). In all studied groups, SNP-induced motor impairment (expressed in %) was lower during the dark than the light phase. Furthermore, there was a non-significant gender-related difference in SNP-induced neuronal toxicity with the males more sensitive than females at every studied PND. The ataxic effects were inversely proportional to the lag time from injection and to the age of animals (with P < 0.05 only between 2 and 8 week old mice). These data indicate that both the administration time and age of the animal significantly affect the neurotoxic effects of SNP.

Differential hypothalamic tyrosine hydroxylase distribution and activation by light in adult mice reared under different light conditions during the suckling period

In mammals, early light experience during a critical period within the first 3 weeks of postnatal development has long-lasting effects on circadian locomotor activity behaviour and neuropeptide expression in the suprachiasmatic nucleus (SCN) of the hypothalamus, site of the principal pacemaker. Dopamine is thought to be involved in the modulation of photic input within the SCN and in tadpoles, the expression of tyrosine hydroxylase (TH), a rate-limiting enzyme in the synthesis of dopamine, in the SCN is altered by previous light history. We thus hypothesised that dopaminergic neurons may be important for the development of the adapted responses to light that we have previously observed. To test this, we raised mice in either constant darkness, 12:12 h light-dark cycles or constant light during the first 3 weeks after birth, and later examined the expression of TH and FOS in the hypothalamus of these mice as adults, both in the dark and after exposure to a light pulse. We found that early light experience affects TH and FOS expression, both baseline levels and in response to a light pulse, in brain areas which are directly connected to the SCN, and are associated with the circadian control of neuroendocrine function. Therefore, our results suggest that the long-lasting alterations induced by early light environment on several hypothalamic nuclei may be relayed through the SCN, and that TH-expressing cells may play a role in conveying/establishing these alterations. These data suggest a role of early light experience in the regulation of future hormonal homeostasis and circadian behaviour.

Mechanisms of Compartmental Purkinje Cell Death and Survival in the Lurcher Mutant Mouse

The Lurcher mutant mouse is characterized by its ataxic gait and loss of cerebellar Purkinje cells and their afferents, granule cells and olivary neurons, during the first weeks of postnatal development. For the 50 years since its discovery, the heterozygous Lurcher mutant has served as an important model system for studying neuron-target interactions in the developing cerebellum and cerebellar function. The identification of the Lurcher (Lc) gene over 10 years ago as a gain-of-function mutation in the δ2 glutamate receptor (GluRδ2) led to extensive studies of cell death mechanisms in the Lc/+ cerebellum. The advantage of this model system is that GluRδ2(+) receptors and GluRδ2(Lc) channels are expressed predominantly in Purkinje cells, making it possible to study the effects of a well-characterized leak current in a well-defined cell type during a critical phase of neuronal development. Yet there is still controversy surrounding the mechanisms of neuronal death in Lc/+ Purkinje cells with competing hypotheses for necrotic, apoptotic, and autophagic cell death pathways as a consequence of the excitotoxic stress caused by the GluRδ2(Lc) leak current. The goal of this review is to summarize recent studies that critically test the role of various cell death pathways in Lc/+ Purkinje cell degeneration with respect to evidence for the molecular heterogeneity of Purkinje cells. We propose that the expression of putative survival factors, such as heat shock proteins, in a subset of cerebellar Purkinje cells may affect cell death pathways and account for the pattern and diverse mechanisms of Lc/+ Purkinje degeneration.

Impaired development of hippocampal mossy fibre synapses in mouse mutants for the presynaptic scaffold protein Bassoon

Bassoon, a protein highly concentrated at the synaptic active zone, is thought to participate in the organization of the cytomatrix at the site of neurotransmitter release. Bassoon is amongst the first proteins to accumulate at newly formed synaptic junctions, raising the question of the functional role of this protein in the early stages of synaptic development. Here we show that the course of synaptic maturation of hippocampal mossy fibre (MF) synapses (glutamatergic synapses with multiple release sites) is markedly altered during the first 2 weeks of postnatal development in mutant mice lacking the central region of Bassoon (Bsn(-/-) mice). At postnatal day 7 (P7), Bsn(-/-) mice display large amplitude MF-EPSCs with decreased paired pulse ratios, an abnormality which may be linked to deficits in the organization of the presynaptic active zone. Surprisingly, 1 week later, decreased MF-EPSCs amplitude is observed in Bsn(-/-) mice, consistent with the inactivation of a subset of synaptic release sites. Finally, at more mature states a decreased posttetanic potentiation is observed at MF-synapses. These results support the notion that Bassoon is important for organizing the presynaptic active zone during the postnatal maturation of glutamatergic synapses.

Early Enriched Environment Promotes Neonatal GABAergic Neurotransmission and Accelerates Synapse Maturation

Environmental stimulation is critical for brain development. Here, we report that natural stimulation through enriched environment (EE) rearing during the first 2 weeks of mouse postnatal development promotes GABAergic neurotransmission and accelerates maturation of GABAergic and glutamatergic synapses. Using whole-cell recordings from CA1 pyramidal neurons in acute hippocampal slices, we found that EE-reared mice exhibited higher amplitude of miniature GABAergic postsynaptic currents (mGPSCs) at 1 week of postnatal development, as well as accelerated transition of GABA action from excitation to inhibition, compared with mice reared under standard housing conditions. This enhanced GABAergic synaptic transmission persisted until the end of the second postnatal week, when GABA mostly acts as an inhibitory neurotransmitter. Consistent with these electrophysiological results, we observed elevated levels of GABA(A) receptors and the K(+)-Cl(-) cotransporter KCC2. Similarly, increased levels of excitatory synaptic components, including NMDA and AMPA receptors and the scaffolding protein PSD95, were detected in synaptosomal fractions from the forebrain/hippocampus of EE-reared mice during the first two postnatal weeks. Functional increase in glutamatergic synaptic transmission, as measured by increased amplitude of miniature and spontaneous EPSCs, was also detected during the second postnatal week. Together, these results demonstrate that early environmental stimulation through EE rearing enhances early postnatal GABAergic neurotransmission, which is known to play important trophic functions in many aspects of neural development.

Developmental sensory experience balances cortical excitation and inhibition

Early in life, neural circuits are highly susceptible to outside influences. The organization of primary auditory cortex (AI) in particular is governed by acoustic experience during the critical period, an epoch near the beginning of postnatal development throughout which cortical synapses and networks are especially plastic1-8. This neonatal sensitivity to the pattern of sensory inputs is believed to be essential for constructing stable and adequately adapted representations of the auditory world and for the acquisition of language skills by children5,9,10. One important principle of synaptic organization in mature brains is the balance between excitation and inhibition, which controls receptive field structure and spatiotemporal flow of neural activity11-15, but it is unknown how and when this excitatory-inhibitory balance is initially established and calibrated. Here we used whole-cell recording to determine the processes underlying the development of synaptic receptive fields in rat AI. We found that, immediately after hearing onset, sensory-evoked excitatory and inhibitory responses were equally strong, although inhibition was less stimulus-selective and mismatched with excitation. However, during the third week of postnatal development, excitation and inhibition became highly correlated. Patterned sensory stimulation drove coordinated synaptic changes across receptive fields, rapidly improved excitatory-inhibitory coupling, and prevented further exposure-induced modifications. Thus the pace of cortical synaptic receptive field development is set by progressive, experience-dependent refinement of intracortical inhibition.

Glial Glucokinase Expression in Adult and Post-Natal Development of the Hypothalamic Region

It has recently been proposed that hypothalamic glial cells sense glucose levels and release lactate as a signal to activate adjacent neurons. GK (glucokinase), the hexokinase involved in glucose sensing in pancreatic β-cells, is also expressed in the hypothalamus. However, it has not been clearly determined if glial and/or neuronal cells express this protein. Interestingly, tanycytes, the glia that cover the ventricular walls of the hypothalamus, are in contact with CSF (cerebrospinal fluid), the capillaries of the arcuate nucleus and adjacent neurons; this would be expected for a system that can detect and communicate changes in glucose concentration. Here, we demonstrated by Western-blot analysis, QRT–PCR [quantitative RT–PCR (reverse transcription–PCR)] and in situ hybridization that GK is expressed in tanycytes. Confocal microscopy and immunoultrastructural analysis revealed that GK is localized in the nucleus and cytoplasm of β1-tanycytes. Furthermore, GK expression increased in these cells during the second week of post-natal development. Based on this evidence, we propose that tanycytes mediate, at least in part, the mechanism by which the hypothalamus detects changes in glucose concentrations.

Postnatal Development and Functional Adaptations of the Melanopsin Photoreceptive System in the Albino Mouse Retina

To study the melanopsin system of the albino CD1 mouse retina during postnatal development. Pups were kept under different ambient conditions: light/dark (LD) cycles, constant light (LL), constant darkness (DD), LL followed by LD, and DD followed by LL. Using immunohistochemistry, melanopsin-expressing cells were classified as M1 or M2 according to the location of their somata and dendritic processes and were counted. Under LD cycles an increase in the number of immunoreactive cells was observed within the first week of postnatal development. When mice were maintained in DD, the increase in the number of immunopositive cells detected was significantly higher than that in LD. On the contrary, when mice were exposed to LL within the same period, no increase was detected. To determine whether the effect of LL during the early postnatal period was reversible, the authors studied animals born in LL and subsequently maintained under LD cycles. After 3 days in LD, these animals showed a significant increase in melanopsin cell number. However, after 1 month in LD, the number was similar to that of the LD controls. Surprisingly, when mice born in DD were exposed to LL, no decrease was detected, though the immunostaining was of low intensity. The amount of melanopsin protein per cell varies, depending on ambient light conditions. Periods of darkness or, more likely, the sequence of light and dark periods occurring under the daily cycles might be necessary for the normal development of the melanopsin system.