The 13th J. A. F. Stevenson memorial lecture. Sexual differentiation of the brain: possible mechanisms and implications

The mammalian brain appears to be inherently feminine and the action of testicular hormones during development is necessary for the differentiation of the masculine brain both in terms of functional potential and actual structure. Experimental evidence for this statement is reviewed in this discussion. Recent discoveries of marked structural sex differences in the central nervous system, such as the sexually dimorphic nucleus of the preoptic area in the rat, offer model systems to investigate potential mechanisms by which gonadal hormones permanently modify neuronal differentiation. Although effects of these steroids on neurogenesis and neuronal migration and specification have not been conclusively eliminated, it is currently believed, but not proven, that the principle mechanism of steroid action is to maintain neuronal survival during a period of neuronal death. The structural models of the sexual differentiation of the central nervous system also provide the opportunity to identify sex differences in neurochemical distribution. Two examples in the rat brain are presented: the distribution of serotonin-immunoreactive fibers in the medial preoptic nucleus and of tyrosine hydroxylase-immunoreactive fibers and cells in the anteroventral periventricular nucleus. It is likely that sexual dimorphisms will be found to be characteristic of many neural and neurochemical systems. The final section of this review raises the possibility that the brain of the adult may, in response to steroid action, be morphologically plastic, and considers briefly the likelihood that the brain of the human species is also influenced during development by the hormonal environment.     

Kinetics and in vitro origin of the temperature-dependent transition of the estrogen receptor monomer

Partitioning of estrogen receptors in aqueous two-phase polymer systems has provided the basis for a detailed kinetic analysis of the effects of temperature on estrogen receptor (ER) structure in vitro. Exposure to temperatures of 0-30 degrees C increased the rate of change in ER partition coefficients by up to 100-fold but did not affect the final extent of the process. The temperature-dependent change in ER partition coefficients was characterized by a linear Arrhenius plot and an activation energy of 25 kcal/mol. The rate of the temperature-dependent ER transition (28 degrees C) was found to be unaffected by greater than 50-fold changes in receptor concentration, which indicates that the temperature-dependent change in partition coefficients reflects a first-order process. The partition coefficients of heated ER were unaffected by subsequent 18-h incubations at 0 degree C, indicating that the temperature-dependent ER transition is irreversible in vitro. Direct heating of the unoccupied ER resulted in both a change in ER partition coefficients and a loss of ER binding sites. The temperature-dependent change in unoccupied ER partition coefficients was complete within 30 min at 28 degrees C and yielded a first-order rate constant that was the same as that obtained for heating the receptor-estradiol complex at 28 degrees C. In contrast, the loss of unoccupied ER binding sites that occurred during 28 degrees C incubations did not reach completion after 150 min of heating and was found to behave as a second-order process.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serological recognition of HLA-DR allodeterminant corresponding to DNA sequence involved in gene conversion

HLA class 11 molecules were isolated from mouse L cells transfected with a DR gene and an allele, 52a, of locus DR III from an HLA-homozygous cell line, AVL, of the DR3 haplotype. The isolated molecules were found to possess a new allospecificity, named TR81. This specificity behaved allelic to the previously described DR III locus. The TR81 specificity was also present on the DR I gene product of the DR3 haplotype. The nucleotide sequence of the gene encoding TR81 differs from TR81-negative DR genes of the DRw52 family in only two codons, both located in the regions known to be involved in a gene conversion event. Consequently, the following conclusions can be formulated. (a) TR81 is a bi-locus specificity and allelic to TR22 only in its DR III locus localization. (b) The TR81 specificity is the phenotypic counterpart of the gene conversion event which led to the generation of the DR I gene of the DR3 haplotype. (c) One or both individual amino acid substitutions in the first domain of the DR chain are responsible for the TR81 allospecificity. (d) Since TR81 is expressed on the DR I chain of the DR3 haplotype, it is possible that TR81 and DR3 represent the same serological specificity.     

Chromosomal assignment of 46 brain cDNAs.

Expressed sequence tags (ESTs) have been obtained from several hundred brain cDNAs as an initial effort to characterize expressed brain genes. These ESTs will become tools for human genome mapping and they will also provide candidate causative genes for inherited disorders affecting the central nervous system. We have developed a procedure for the rapid chromosomal assignment of these ESTs: cDNA sequences are first analyzed by a computer program to determine regions likely not to be interrupted by introns in the genomic DNA. A pair of oligonucleotide primers is then designed to amplify this region by the polymerase chain reaction using DNA template from human-rodent somatic cell hybrid chromosomal panels. The chromosomal assignment of the cDNA is determined by studying the segregation of the amplified products in these panels. In this paper we describe the mapping of 46 brain ESTs, as well as observations on the amplification of rodent sequences.     

Proestrous hormonal changes preceding the onset of ovulatory failure in lightly androgenized female rats

Proestrous hormonal profiles were characterized in lightly androgenized female rats prior to the onset of the delayed anovulatory syndrome (DAS). In these females, ovulatory failure and persistent vaginal estrus (PVE) occur at a very early age. Female Sprague-Dawley rats were injected with 10 micrograms testosterone propionate (TP) on postnatal Day 5. Control rats were untreated. All animals were weaned at 21 days of age, and following the onset of puberty, estrous cyclicity was monitored by vaginal smear. Rats showing regular 4-day cycles were used. Between 50-70 days of age, intra-atrial cannulae were implanted on a morning of proestrus (0700-0900 h) and blood was sampled at 2-h intervals from 1000 to 2000 h. Additional samples were taken at 0.5-h intervals from 1600 to 1800 h. Plasma was assayed for luteinizing hormone (LH), follicle-stimulating hormone (FSH) and progesterone (P) by radioimmunoassay (RIA). All animals were monitored for the onset of PVE or other alterations in estrous cyclicity. Females treated neonatally with TP that subsequently showed PVE by 150 days of age (PRE DAS) displayed a reduced peak amplitude (P less than 0.01) and delay in onset (1600 vs. 1400 h) of LH but not FSH secretion, when compared to controls. Females treated neonatally with TP that did not enter PVE by 150 days of age (No DAS) also showed a delayed rise in LH when compared to controls. However, the amplitude of LH secretion was not different from controls or PRE DAS females.(ABSTRACT TRUNCATED AT 250 WORDS)     

Unilateral lesions of the hamster suprachiasmatic nuclei: evidence for redundant control of circadian rhythms

Summary Several properties of vertebrate circadian rhythms can be attributed to the behavior of an underlying pacemaker system which is composed of two separate but mutually interacting circadian oscillators. As originally formulated, the model for such a pacemaker system proposed that two oscillators or populations of oscillators have different properties, specifically in their responses to light (Pittendrigh 1974; Pittendrigh and Daan 1976b). We have tested the proposition that the right and left suprachiasmatic nuclei (SCN) of the golden hamster contribute in different ways to the regulation of circadian rhythmicity by measuring the wheel-running activity rhythms of hamsters with lesions to either the right or left SCN. Although effects of unilateral or other partial SCN lesions on pacemaker properties were observed, these effects were not different in hamsters receiving right- or left-side lesions. More specifically: (1) free-running period () in constant light was shorter in lesioned hamsters irrespective of the side lesioned (Fig. 3a), and the total amount of SCN destruction was found to correlate with (Fig. 4). (2) Phase-angle difference () of some lesioned hamsters (both right- and left-side) during entrainment to LD, 1410 was significantly more positive than that of controls (Fig. 3b). (3) The rate of phase-shift following a shift of the light/dark cycle was not different in hamsters with right- or left-side lesions (Fig. 3c). And (4) the simultaneous expression of different circadian periods, similar to splitting, was observed in hamsters with unilateral lesions (Fig. 5). It is concluded that the right and left SCN are similar in their contributions to the control of circadian rhythmicity and that there is as yet no evidence for the permanent loss of multioscillator properties resulting from the destruction of only one of the two SCN.Abbreviations SCN suprachiasmatic nuclei or nucleus - LD light/dark cycle - LL constant light - DD constant dark - circadian period - activity time - rest time - phase angle - phase-angle difference - SD standard deviation - SE standard error - ANOVA analysis of variance     

Purification and properties of microsomal UDP-glucuronosyltransferase from rat liver.

p-Nitrophenol conjugating activity associated with liver microsomal UDP-glucuronosyltransferase (EC 2.4.1.17) was purified 150- to 200-fold from cell-free homogenates. The purification scheme included solubilization with the nonionic detergent Lubrol WX, anion exchange chromatography at pH 6.0 and 7.5, and affinity chromatography with UDP-hexanolamine Sepharose 4B. The enzyme purified as a phospholipid-protein complex and was shown to consist of a single polypeptide chain of molecular weight 59,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Amino acid analysis indicated approximately 531 mol of amino acids/59,000 g of enzyme and a molar ratio of nonpolar to polar residues of 1.08. During fractionation, the enzyme displayed instability with such steps as gel filtration, dialysis, or ultrafiltration of dilute samples; however, upon adsorption to ion exchange resins or storage in concentrated form, the enzyme was reasonably stable. The active lipoprotein complex showed both size and charge heterogeneity as judged by gel filtration and electrofocusing. Three forms of the enzyme resolved by isoelectric focusing had isoelectric points which averaged pH 6.68, 6.56, and 6.31. Polypeptide compositions of these electrophoretically distinct phospholipid protein complexes were indistinguishable on the basis of sodium dodecyl sulfate-polyacryl-amide gel electrophoresis, suggesting that the charge heterogeneity may be the result of differences in the phospholipid content of the lipoprotein complex.