Expression of the alpha and beta subunits of Ca2+/calmodulin kinase II in the cerebellum of jaundiced Gunn rats during development: a quantitative light microscopic analysis.

The homozygous (jj) jaundiced Gunn rat model for hyperbilirubinemia displays pronounced cerebellar hypoplasia. To examine the cellular mechanisms involved in bilirubin toxicity, this study focused on the effect of hyperbilirubinemia on calcium/calmodulin-dependent kinase II (CaM kinase II). CaM kinase II is a neuronally enriched enzyme which performs several important functions. Immunohistochemical analysis of alternating serial sections were performed using monoclonal antibodies for the alpha and beta subunits of CaM kinase II. Measurements were made of the total numbers of stained cells in each of the deep cerebellar nuclei and of Purkinje and granule cell densities in cerebellar lobules II, VI, and IX. The beta subunit was present in Purkinje cells and deep cerebellar nuclei of both groups at all ages, but only granule cells which had migrated through the Purkinje cell layer showed staining for beta subunit; external granule cells were completely negative. Many Purkinje cells had degenerated in the older animals, and the percent of granule cells stained for beta subunit was significantly reduced. The alpha subunit was found exclusively in Purkinje cells, although its appearance was delayed in the jaundiced animals. Sulfadimethoxine was administered to some jj rats 24 h or 15 days prior to sacrifice to increase brain bilirubin concentration. Results showed that bilirubin exposure modulated both alpha and beta CaM kinase II subunit expression in selective neuronal populations, but sulfadimethoxine had no acute effect on enzyme immunoreactivity. Thus, developmental expression of the alpha and beta subunits of CaM kinase II was affected by chronic bilirubin exposure during early postnatal development of jaundiced Gunn rats.

Development of cerebellar hypoplasia in jaundiced Gunn rats: a quantitative light microscopic analysis.

The homozygous (ii) Gunn rat provides a model for hyperbilirubinemia which includes prominent cerebellar hypoplasia. Development of the Gunn rat cerebellum was examined with and without the additional effects of elevating brain bilirubin concentration to still higher levels via sulfadimethoxine (sulfa) administration. Homozygous (jj) Gunn rats and heterozygous (Nj) littermate controls (n = 32 each) were given 100 mg/kg sulfa or saline at postnatal days 3, 7, 17, and 30, and most were sacrificed 24 h later (n = 4 for each genotype at each age). Cerebellar volume, total volume and cell number for each deep cerebellar nucleus, densities for Purkinje and granule cells in the cerebellar cortex of lobules II, VI and IX, and the density of vacuolated Purkinje cells were all measured quantitatively. Cytoplasmic vacuolation provided an indication of bilirubin toxicity and was never observed in the Nj control rats. Vacuolated Purkinje cells were first observed in jj-saline rats at 18 days and were found only in the more anterior lobules of the cerebellum (II and VI). By contrast, vacuolated Purkinje cells were observed in jj-sulfa rats at both 4 and 8 days, but only in the most posterior cerebellar lobule (IX). In all older jj rats, the decline in vacuolation was accompanied by significant necrosis and resorption of the Purkinje cells in the anterior lobules. Since the Purkinje cells in the posterior lobules are the first to differentiate in the cerebellum and are resistant to bilirubin toxicity in jj-saline rats, the results support the presence of a critical period when elevated brain bilirubin may be most toxic to neuronal development. The findings suggest that neurons undergoing differentiation at the time of bilirubin exposure are most susceptible to cell death, while cells that are slightly more or slightly less mature may show only transient changes.

Differential effects of gentamicin on the distribution of cochlear function in albino and pigmented guinea pigs.

It has been suggested that the high affinity of melanin pigment for aminoglycoside antibiotics may cause these drugs to bind preferentially to the pigmented inner ear, producing greater ototoxicity than in the amelanotic albino cochlea. However, evidence of greater ototoxicity in albinos has led to the hypothesis that melanin inhibits the toxicity of these drugs in the pigmented inner ear. On the other hand, ototoxicity in the pigmented animals may simply be delayed relative to the albinos, only to become equal or even more severe with time. The present study was conducted to determine whether a relatively low dose of gentamicin (68.5 mg/kg) would produce differential ototoxicity between albino and pigmented guinea pigs which would persist long after drug exposure had stopped. Nine pigmented and eight albino guinea pigs were given gentamicin sulfate for 14 consecutive days, and were then allowed a two-month recovery period before cochlear analysis; 11 pairs of saline-injected or untreated albino and pigmented guinea pigs served as controls. The results showed that the gentamicin-treated albinos had significantly elevated thresholds for the compound action potential from the auditory nerve (CAP), and significantly lower endocochlear potentials (EP) and cochlear microphonic (CM) input-output voltage functions when compared to their respective controls, or to either group of pigmented guinea pigs. The CAP in drug-treated pigmented animals did not differ significantly from controls, and the differences in EP and CM were marginally significant. The results indicate that the pigmented cochlea is less susceptible to gentamicin than the albino cochlea, and support the hypothesis that melanin may inhibit aminoglycoside ototoxicity in the pigmented inner ear.

Ongoing proliferation of melanocytes in the stria vascularis of adult guinea pigs.

The intermediate cells of the stria vascularis are melanocytes derived from the neural crest. These internalized pigment cells have been thought to be a static population, distinct from those found in the skin. However, this investigation demonstrates that the melanocytes of the adult stria vascularis undergo continuous replication. Cell proliferation was studied using [3H]-thymidine autoradiography and bromodeoxyuridine (BrdU) immunohistochemistry. Single or multiple injections of [3H]-thymidine within a six hour period labeled a mean of 9 intermediate cells. In pigmented guinea pigs, single daily injections of [3H]-thymidine for 2 or 4 days labeled a mean of 24 and 69 intermediate cells, respectively; Pigmented guinea pigs given BrdU once daily for 2 or 4 days labeled a mean of 38 and 75 intermediate cells. By contrast, albino littermates also given BrdU averaged only 23 and 42 labeled intermediate cells in the same 2 and 4 day experiments. The mean number of proliferating cells/mm of stria per 24 h was 1.54 in the pigmented animals and 0.88 in the albinos. Both the total number and density of labeled intermediate cells were significantly smaller in the albino than the pigmented guinea pigs. These results demonstrate that the melanocytes in the stria vascularis undergo continuous baseline mitosis, and at a rate comparable to the melanocytes of the skin. This surprising similarity promotes the speculation that the proliferative rate of the strial melanocytes may be influenced by some of the same factors known to affect replication and pigment production in the skin.

Auditory brainstem responses in cats with Chediak-Higashi syndrome.

Auditory brainstem responses ABRs were recorded in cats with Chediak-Higashi syndrome using monaural stimulation. The components appearing between 1 and 3 ms after stimulus onset were greatly attenuated in the ABRs recorded using a reference contralateral to the stimulated ear. These data suggest that abnormalities exist in the brainstem auditory pathway in the region of the superior olivary complex in cats with Chediak-Higashi syndrome.

Pigment-dependent differences in the stria vascularis of albino and pigmented guinea pigs and rats.

Functional models of the stria vascularis (SV) have ascribed roles for the marginal and basal cells, but not for the intermediate cells, which remain poorly understood. Intermediate cells have been identified as melanocytes, which produce melanin in most pigmented animals including humans. The relationship of melanin to intermediate cell function may be addressed through comparisons with the albino inner ear. Albinos have a normal distribution of melanocytes that are unable to synthesize melanin pigment. In the present study, the SV was compared between albino and pigmented littermates in both the guinea pig and the rat. Photomicrographic montages of the SV were analyzed from each of 7 cochlear regions in the guinea pig and 5 regions in the rat. Stereological procedures were used to determine the volume density (Vv) for each of the three main cell types in the stria, the surface density (Sv) of the marginal cells, and to derive estimates of absolute cell volume and surface area. In the guinea pig, comparisons between pigment groups showed that marginal cell Vv was larger across cochlear turns in the albinos, while intermediate cell Vv was smaller. Intermediate cell cytoplasmic and total cell volumes were smaller in the albino guinea pigs; however, marginal cell Sv and absolute area were larger. In the rat, intermediate cell Vv was alos smaller across cochlear turns in the albinos. Similarly, intermediate cell cytoplasmic and total cell volumes were smaller in the albinos, while marginal cell total surface area per radial cross-section of the SV was larger. These results demonstrate that amelanotic melanocytes occupy significantly less volume than do pigmented melanocytes, and suggest that melanin may influence the structure and function of the SV.

Turn-specific differences in the endocochlear potential between albino and pigmented guinea pigs.

Recent findings indicate that structural differences exist in the stria vascularis (SV) between albino and pigmented guinea pigs. In the higher cochlear turns, volume density for marginal cells in the albino SV is abnormally large, while that for intermediate cells (melanocytes) is abnormally small. These anatomical variations suggest that functional differences between albino and pigmented inner ears also may be found. To examine this possibility, four strains of guinea pigs were studied, consisting of Hartley albino (N = 9) and NIH outbred pigmented (N = 15) guinea pigs, as well as albino (N = 11) and pigmented (N = 15) guinea pig siblings born to mixed litters. Tracheotomy and carotid artery cannulation were performed. Animals were mechanically ventilated, with periodic samples drawn for arterial blood gas analysis. Blood pressure, heart rate and rectal temperature were monitored. Compound action potentials were measured first to assess cochlear viability. Positive endocochlear potentials (+EP) then were recorded, beginning with the fourth turn, followed by the first, second and third turns. Results showed that the +EP in albinos remained relatively constant across cochlear turns, but decreased significantly from base to apex in the pigmented inner ears. Across all animals, mean +EPs (mV +/- S.E.M.) for turns 1-4 in albinos were: 72.5 (2.5), 68.7 (2.3), 59.2 (2.7), 68.1 (3.3); pigmented values were: 72.9 (2.9), 66.9 (2.6), 53.8 (3.0), 57.0 (2.7). One-way ANOVAs did not show a significant difference in albino +EPs between any of the cochlear turns, but did indicate a highly significant difference between turns in the pigmented inner ears (P < 0.000004). Post hoc comparisons demonstrated +EPs in turns 3 and 4 were smaller than in turn 1. Since turn 3 was recorded last in these experiments, and was reduced in value relative to turn 4 in both groups, it is likely that cochlear deterioration contributed to this result more than any other factor. These results, combined with previous anatomical data, indicate that a diminution of melanocyte cell volume in the albino SV is accompanied by an increase in marginal cell volume density and larger +EPs in the higher cochlear turns, at least at resting levels.

Brainstem auditory evoked potentials correlate with morphological changes in Gunn rat pups.

The relationship of brainstem structure and function in bilirubin encephalopathy is incompletely understood. The present experiments compare quantitative measures of brainstem structures with brainstem auditory evoked potentials (BAEPs) in infant jaundiced (jj) and nonjaundiced (Nj) Gunn rats. Ten jj's from 4 litters were injected with sulfadimethoxine at 11-12 days of age to raise their brain bilirubin concentration. Littermate controls were jj's given saline, and Nj's given sulfadimethoxine or saline. At 15-17 days of age BAEPs were recorded, and rats were prepared for histological examination, as was reported in the previous paper (Conlee and Shapiro, 1991). Significant differences between groups were seen for BAEP wave I latency (P = 0.002). I-II interwave interval (P = 0.001), and amplitudes of waves I, II, III, and IV (each P less than 0.0005) due to increased latencies and decreased amplitudes in the jj-sulfa group. Animals with the most severe BAEP abnormalities had the most severe histological abnormalities. Cochlear nucleus volume had a positive linear correlation with the amplitude of BAEP waves I, II, and IV, and an inverse correlation with wave I latency and I-II interwave interval (P less than or equal to 0.001). The highest correlations were BAEP I-II interwave interval and amplitude of waves I and II with cochlear nucleus volume (r = -0.78, 0.71 and 0.70, respectively, P less than 0.0005).(ABSTRACT TRUNCATED AT 250 WORDS)

Morphological changes in the cochlear nucleus and nucleus of the trapezoid body in Gunn rat pups.

Mechanisms underlying bilirubin encephalopathy and hearing loss remain poorly understood, including the way bilirubin enters the nervous system and how bilirubin accumulates in circumscribed regions of the brain. The present experiments examined the auditory brainstem in heterozygous (Nj) and homozygous (jj) Gunn rats at an age when serum bilirubin levels were highest, and after brain bilirubin concentration was artificially raised by sulfadimethoxine administration. In four litters of 11-12 days old Gunn rats, Nj and jj littermates received a single intraperitoneal injection of sulfadimethoxine (100 mg/kg) or a comparable volume of saline. At 16-17 days of age, brainstem auditory evoked potentials were recorded to assess the severity of bilirubin toxicity in the Nj and jj animals. Following the recordings, each animal was perfusion-fixed and frozen sections of the brainstem were cut in the transverse plane from medullary through mesencephalic levels. Sections were mounted on slides, stained with thionin and coded to avoid observer bias. Quantitative analysis revealed no differences between saline and sulfa-treated Nj rats for cochlear nucleus volume, or for cell size in the cochlear nucleus or superior olive. In the sulfa-treated jj rats, cochlear nucleus volume, and cross-sectional areas of spherical cells in the anteroventral cochlear nucleus and principal cells in the nucleus of the trapezoid body, were all significantly smaller than in the combined groups of Nj animals. The affected areas in the cochlear nucleus and superior olive are innervated by large axosomatic end-bulbs of Held or calyceal endings, and were associated with bilirubin staining of glia in the most severely jaundiced jj sulfa-treated rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Turn-specific and pigment-dependent differences in the stria vascularis of normal and gentamicin-treated albino and pigmented guinea pigs.

The aims of the present study were to determine which structures in the stria vascularis (SV) may depend upon the presence of pigmented melanocytes both for normal morphology and for the expression of gentamicin ototoxicity in the inner ear. These pigment-dependent influences were inferred through comparisons of the SV in pigmented guinea pigs and in albinos containing nonpigmented melanocytes. Results were obtained from 6 albino and 8 pigmented guinea pigs given gentamicin, and from 3 albino and 3 pigmented control animals not receiving the drug. One-month old animals received gentamicin daily (100 mg/kg) for 14 days and recovered for an additional 14 days before being prepared for electron microscopy. The SV from each of the 4 cochlear turns was analyzed using stereological point counting procedures. In control animals, differences were found in the higher cochlear turns, where volume density for the marginal cells in albinos was abnormally large (turns 3 and 4), while the volume density for intermediate cells (melanocytes) was abnormally small (turn 3). Cell volume estimates for the intermediate cells were significantly smaller in the albino than pigmented control animals in the higher cochlear turns, indicating that functional abnormalities may be found in the albino cochlea. In animals exposed to gentamicin, marginal cell volume density was reduced significantly in turn 4 of albinos, but not in any region of the pigmented inner ears. Radial area of SV and estimates of the absolute volumes for marginal cells in albinos given gentamicin also were significantly reduced in turn 1 compared to their controls; such differences were not observed in the pigmented animals. The results indicate that marginal cell size is significantly reduced in albino but not pigmented animals 14 days after gentamicin exposure, and further suggest a role of pigmented melanocytes in ameliorating gentamicin-induced cochlear damage.

Differential susceptibility to gentamicin ototoxicity between albino and pigmented guinea pigs.

The known chemical affinity of melanin pigment for aminoglycoside antibiotics has led to the suggestion that higher concentrations of these drugs will bind to the pigmented inner ear and produce greater ototoxicity compared to the nonpigmented albino cochlea. Although this has provided a compelling hypothesis, results from the few investigations to address this question have been equivocal. In the present study, cochlear microphonic (CM) thresholds were recorded from albino and pigmented guinea pigs both before and two weeks after exposure for 14 consecutive days to 100 mg/Kg gentamicin. Cochleae were dissected and half-turn segments prepared for surface examination of the organ of Corti. After gentamicin exposure, threshold shifts averaged a statistically reliable 33 dB in albinos and 19 dB for the pigmented animals. Anatomical studies revealed a significant 44% mean outer hair cell loss in albinos compared to a 21% loss in the pigmented inner ears. The results showed that albinos display greater ototoxicity from gentamicin than do pigmented guinea pigs. Aminoglycosides are known to exert toxicity through interaction with polyphosphoinositides found in high concentrations in the inner ear. Cochleae in both albino and pigmented animals appear to possess significant phospholipid concentrations and bind toxic levels of these drugs independent of inner ear pigment content. However, evidence showing that melanin can inhibit aminoglycoside activity in vitro suggests that, once these drugs bind to pigmented tissue, they may undergo inactivation in a manner unavailable to the nonpigmented albino cochlea. The present results are consistent with the possibility that cochlear melanin may inhibit gentamicin activity in vivo and decrease the severity of aminoglycoside ototoxicity in the pigmented inner ear.

Comparative anatomy of melanin pigment in the stria vascularis. Evidence for a distinction between melanocytes and intermediate cells in the cat.

Although Corti in 1851 first described the presence of cochlear pigmentation in the stria vascularis (SV) of "very old" cats, modern studies have failed to find pigment consistently in the feline stria. While the variable presence of pigment in the feline SV would appear to contrast with this structure's uniform pigmentation in other mammalian species, variability in both the distribution and abundance of inner ear pigment has rarely been studied in any species. In the present study, the SV was examined light microscopically in sectioned material or whole-mounts from pigmented and albino animals of 5 species, including the cat, guinea pig, rabbit, ferret and mouse. In these species, the SV of each pigmented animal contained varying amounts of melanin pigment and none was found in the albino inner ear. Pigmented guinea pigs contained the most uniformly dense and least variable distribution of strial melanin, followed by the rabbit, mouse, ferret and cat. Several species also displayed more strial pigment apically and less basally. In cats, pigmented cells were principally located adjacent to the strial capillaries. Ultrastructural studies of the stria in pigmented cats revealed that these perivascular cells frequently contained an abundance of pigmented organelles and other structural features which allowed them to be distinguished from intermediate cells.

Effects of aging on normal hearing loss and noise-induced threshold shift in albino and pigmented guinea pigs.

In a previous investigation into noise-induced hearing loss by comparing 2-month-old albino with pigmented guinea pigs, albinos displayed significantly greater shifts in cochlear microphonic (CM) threshold and less recovery than the pigmented animals 7 days after noise exposure. The present study compared the responses of 14-month-old albino and pigmented guinea pigs to the same noise parameters used previously. Thresholds for the first detectable elicitation of CM for three pure tones were recorded prior to, at 90 min and at 7 days after a 45-min exposure to 126 dB broadband noise. Before exposure to noise, thresholds for pigmented guinea pigs were 24 dB higher than those in the albinos. Following noise exposure, the pigmented animals showed less than half the amount of threshold shift displayed by the albinos. This change ws attributed to the higher pre-exposure thresholds in the pigmented guinea pigs. Converging lines of evidence suggest that cochlear pigmentation may have both protective and toxic influences on the inner ear.

Origin of ascending auditory projections to the nucleus mesencephalicus lateralis pars dorsalis in the chicken.

Ascending auditory projections to the nucleus mesencephalicus lateralis pars dorsalis (MLd) were studied in white Leghorn chickens by means of unilateral injections of horseradish peroxidase into the MLd and by injections of tritiated leucine into nucleus angularis or the combined nucleus magnocellularis and nucleus laminaris. The experiments showed that nucleus angularis sends an extensive projection to the contralateral MLd and a smaller projection to the rostral pole of the ipsilateral MLd; the lagenar region contributes to these bilateral connections. Nucleus angularis also projects bilaterally to the superior olive and nucleus ventralis lemnisci lateralis and to the contralateral nucleus lemnisci lateralis pars ventralis and dorsal nucleus of the lateral lemniscus. Projections from nucleus laminaris were demonstrated to the ipsilateral superior olive, to the contralateral lemniscal nuclei and a small medial region in MLd bilaterally; the contralateral projection is much denser than the ipsilateral one. Other nuclei having ascending connections with MLd include the contralateral superior olive, the ipsilateral nucleus lemnisci lateralis pars ventralis, the contralateral nucleus ventralis lemnisci lateralis and the contralateral MLd. The ipsilateral superior olive and nucleus ventralis lemnisci lateralis also project to MLd but much more sparsely than in their contralateral projection. Although several of these findings correspond with auditory connections previously shown in the pigeon brainstem, they differ fundamentally in that we find both nucleus angularis and nucleus laminaris projecting to different areas of the MLd on both sides of the brain. In particular, our observation that the cochlear nucleus has bilateral connections with MLd demonstrates an important avian similarity with the brainstem auditory pathways of other terrestrial vertebrates.

Differential susceptibility to noise-induced permanent threshold shift between albino and pigmented guinea pigs.

Evidence that reduced levels of cochlear melanin are associated with increased auditory sensitivity, increased levels of auditory fatigue and an increased susceptibility to noise-induced hearing loss led us to investigate the effects of noise exposure on the cochlear microphonic (CM) in albino and pigmented English shorthair guinea pigs. CMs were recorded from the round window prior to and at 90 min and 7 days after exposure to 45 min of 126 dB noise. Thresholds for the first detectable elicitation of the CM for four pure tones were determined and the output voltage of each cochlea was measured in 10 dB steps through intensity levels which produced a maximum voltage amplitude in the CM and voltage rollover. This analysis demonstrated that: albino guinea pigs displayed significantly lower auditory thresholds than did pigmented animals before exposure to noise; thresholds were elevated to comparable levels in both groups 90 min after noise exposure; pigmented guinea pigs showed a reliable recovery in CM thresholds 7 days after exposure to noise while thresholds in the albinos remained elevated to the same degree at both 90 min and 7 days after noise; 90 min after noise exposure, the maximum voltage output of albino cochleas was significantly less than that recorded from the cochleas of the pigmented guinea pigs. These results demonstrate that albino guinea pigs are more susceptible to the ototoxic effects of high intensity noise than pigmented guinea pigs. Converging evidence indicates that some aspects of cochlear function involve melanin pigment and that its absence may produce auditory abnormalities. Reduced melanin pigmentation may also contribute to such phenomena as noise-induced threshold shifts and individual differences in noise-induced hearing loss.

Reduced neuronal size and dendritic length in the medial superior olivary nucleus of albino rabbits.

We have previously demonstrated that circumscribed structural and functional abnormalities exist in the brainstem auditory system of albino cats. Anomalies in the auditory brainstem evoked response of albino cats were correlated with anatomical defects in the medial superior olivary nucleus (MSO) of the same animals. To examine whether a similar syndrome is present in other albino mammals, we studied the MSO of albino and pigmented rabbits using both Nissl-stained and Golgi-impregnated material. Neurons in the MSO of the albinos were significantly smaller (24%) than those in the pigmented rabbits and there was no overlap in the size distributions between the two groups. Neurons in the abducens nucleus of the albinos were also 14% smaller than in the pigmented rabbits, but this difference was not statistically reliable. The broad overlap in the distributions of neuronal size in the abducens nucleus between groups indicated that not all cells in the albino brainstem are significantly smaller than normal. In the Golgi-impregnated material, the mean total dendritic length for the 'marginal' cell type in the MSO was 39% shorter in albinos than in the pigmented animals. The branching density of dendrites was also significantly reduced in the albinos. Mean total dendritic length for cerebellar granule cells was a statistically insignificant 6% longer in the albinos, demonstrating that dendritic structure is not uniformly affected in all regions of the albino brain. The demonstration of similar anatomical differences in albino rabbits and cats indicates that whatever process produces these effects is not species-specific and may be common to the albinos of other mammalian species. The evidence that the amount of cochlear melanin may be related to differences in auditory function further suggests that the differences in the MSO of the albinos may ultimately be related to absence of inner ear pigmentation and not to other gene effects.

Auditory brainstem anomalies in albino cats: II. Neuronal atrophy in the superior olive.

In a previous paper (Brain Res., 260:1-9, 1983) we reported that albino cats show abnormal auditory brainstem evoked responses that appear to arise from structural defects in or near the superior olivary complex. In the present study, neuronal cross-sectional area in brainstem nuclei was compared in albino and normally pigmented adult cats. The albinos were true tyrosinase-negative (cc) and should not be confused with the deaf white cat (W); the albinos are not deaf. Neurons in the medial superior olivary nucleus (MSO) of albinos were, on average, 41% smaller than in pigmented animals; there was no overlap in the neuronal size distributions for the two groups of animals. Cell size in the lateral superior olive, medial nucleus of the trapezoid body, ventral nucleus of the lateral lemniscus, anteroventral cochlear nucleus, dorsal cochlear nucleus, and facial nucleus was also smaller (by 9-21%) in albinos than in pigmented animals but none of these differences was statistically reliable. In the abducens nucleus, neurons were 12% larger in albinos than in pigmented animals, demonstrating that neuronal size in the albinos is not uniformly smaller. Several lines of evidence suggest that the auditory system defects in albinos are related to abnormal pigmentation rather than to other gene effects. It is possible that a subtle pigment-related disruption of inner ear development in albinos results in a central cascade of atrophic changes along the auditory pathway.

Differential rearing affects responsiveness of rats to depressant and convulsant drugs.

The effects of postweaning housing in environmental complexity (EC) or isolation cage (IC) conditions upon the development of anesthesia following sodium pentobarbital administration and upon seizure susceptibility following metrazol administration were studied in separate groups of rats. Male and female EC rats lost the righting reflex and developed unconsciousness more rapidly following sodium pentobarbital injection (105 mg/kg, IP) than did littermate IC rats. Male EC and IC rats did not differ in the CD50 (50% convulsion dosage by log probit analysis) following injection of a range of metrazol doses (20 to 35 mg/kg, IP) when they were kept in a quiet dimly lighted room. However, the addition of stroboscopic light stimulation (1-100 Hz) to the above procedure (doses 17.5 to 30 mg/kg) reduced the CD50 for the IC rats but did not affect the CD50 for the EC rats. The results support the view that brain excitability is lower in animals reared in environments providing higher levels of sensory stimulation.