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L Martini

Publications and source records attributed to L Martini.

At least 109 records · Page 6Linked to original sources
PubMed1995

Characterization of functional opioid delta receptors in a luteinizing hormone-releasing hormone-producing neuronal cell line.

Endogenous opioids participate in the regulation of gonadotropin secretion through an influence on the release of the hypothalamic LHRH. However, it is not clear whether opioids exert a direct effect on LHRH-producing neurons or interfere with other systems able to influence LHRH release. A neuronal LHRH-producing cell line (GT1) developed recently provides a good model to study the mechanisms controlling LHRH release. In the present study, the presence of opioid receptors on a subclone of GT1 cells (GT1-1) has been investigated. A specific and saturable binding of the 3H-labeled nonselective opioid ligand diprenorphine ([3H]DIP) was detected by a receptor binding assay on both intact GT1-1 cells and crude membrane preparations obtained from these cells. Analysis of saturation curves revealed that [3H]DIP apparently binds to a single class of sites with a Kd of 0.2 nM and a binding capacity of 125 fmol/mg protein, corresponding to approximately 20,000 sites/cell. Selective displacement of the binding of [3H]DIP to GT1-1 cells by [D-Ala2,N-Me-Phe4,Gly5-ol]enkephalin, [D-Pen2,D-Pen5]enkephalin (DPDPE), and U50488H, which are selective ligands, respectively, for mu-, delta-, and kappa-receptors, was also evaluated. Only the specific delta-ligand DPDPE produced a significant inhibition of the binding of [3H]DIP. [D-Ala2,N-Me-Phe4,Gly5-ol]Enkephalin and U50488H were totally ineffective. The inhibitory effect of the agonist DPDPE on the binding of [3H]DIP was decreased by the presence of sodium ions, a typical characteristic of the binding of agonists to opioid receptors. Finally, it has been observed that treatment with prostaglandins E1 and E2 produces a dramatic increase in cAMP accumulation in GT1-1 cells, and DPDPE is highly effective in suppressing this effect. On the basis of these results, it is possible to postulate the presence of functional delta-opioid receptors on GT1-1 cells. By extrapolation, one might suggest that endogenous opioids may affect LHRH neurons by two mechanisms: a direct one, acting via delta-receptors, and an indirect one, through the activation of neurons impinging on the LHRH system, which uses mu-receptors.

Animals↗
PubMed1995

Inhibition of luteinizing hormone-releasing hormone secretion by delta-opioid agonists in GT1-1 neuronal cells.

The endogenous opioids play a major role in regulation of the secretion of hypothalamic LHRH. However, it is not clear whether opioids exert a direct effect on LHRH neurons or interfere with other neuronal systems impinging on the cells synthesizing LHRH. The neuronal LHRH-producing cell line GT1 provides a new model to evaluate which signals may directly modify LHRH release. In a previous paper it has been reported that opioid-binding sites of the delta-type are present in a clone of the GT1 cells (GT1-1). In the present study, the possible effects of opioids on the release of LHRH were studied in GT1-1 cells. The results obtained show that only the addition of opioid agonists that bind to delta-receptors brings about a significant inhibition of forskolin- or prostaglandin E2-stimulated LHRH release in GT1-1 cells. The effect of the delta-opioid agonist [D-Pen2,D-Pen5]enkephalin is dose dependent and is reversed by the universal opioid antagonist naltrexone and the delta-specific antagonist naltrindole. No effect of opioid agonists or antagonists was observed in unstimulated cells. These results suggest that opioids may control the release of LHRH also, acting directly on LHRH-producing neurons.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cycloh↗
PubMed1995

Type 1 astrocytes influence luteinizing hormone-releasing hormone release from the hypothalamic cell line GT1-1: is transforming growth factor-beta the principle involved?

The possible existence of a humoral communication between glial cells and LHRH-secreting neurons has been studied using the LHRH-secreting GT1-1 cell line and type 1 astrocytes. Two different designs have been adopted: 1) GT1-1 cells were coincubated with purified cultures of type 1 rat astrocytes, and 2) GT1-1 cells were exposed to the conditioned medium (CM) in which type 1 rat astrocytes had been grown for 24 h. LHRH was measured by RIA in the medium of the GT1-1 cell cultures at different time intervals. The data show that short periods (1, 3, and 6 h) of either coculture or exposure to previously frozen CM significantly increase the release of LHRH from the GT1-1 cells. However, more prolonged times of coculture (e.g. 2 and 5 days) or exposure to CM (e.g. 48 h) induce a significant decrease in the amount of LHRH in the medium. The stimulatory effect on LHRH release appears to be specific for type 1 astrocytes (either cortical or hypothalamic), because neither the CM of oligodendrocytes nor the CM of LNCaP cells (a cell line derived from a human prostatic cancer) possess stimulating activities. Heating the type 1 astrocyte-CM to 100 C for 10 min does not eliminate the ability of the CM to significantly increase the release of LHRH from GT1-1 cells at 1, 3, and 6 h. Because of the opposite effects encountered in the short and long term experiments, it was hypothesized that the CM might contain, in addition to LHRH-releasing principle(s), LHRH-degrading properties. Known amounts of standard LHRH were then added to type 1 astrocyte-CM, either untreated or submitted to heating at 100 C for 10 min. The amount of LHRH added to untreated CM decreases progressively; on the contrary, the amount of LHRH added to heated CM remains unchanged. These results confirm that one or more heat-sensitive enzymes able to degrade LHRH may be present in the type 1 astrocyte-CM. As previously mentioned, the experiments reported so far were performed using type 1 astrocyte-CM that had been kept frozen for various periods of time, before being tested for its LHRH-releasing activity. Surprisingly, fresh CM proves to be inactive, whereas heated CM is effective; this suggests that the factor involved might be activated by the two opposite experimental procedures.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1995

[In vivo experimental research in vascular surgery. Methodology and current Italian law].

Authors underscore the importance of research in vascular surgery progress and approach some basic aspects on the correct conduction of an experimental research in vascular surgery fields and in particular on synthetic prosthesis of different diameter. Some important aspects of the current Italian law on animal experimentation are shown because they could cause new changes either on methodology or animal selection. Finally some experimental models adopted in the department of experimental surgery of the Rizzoli Orthopaedic Institutes on vascular prosthesis are described.

Animal Welfare↗
PubMed1994

First recurrence analysis of 840 cutaneous melanomas: a proposal for a follow-up schedule.

AIMS AND BACKGROUND: A correct follow-up schedule for patients who underwent an excision for stage I cutaneous melanoma might allow the early detection of local and distant metastases. At present, there is no general agreement on follow-up protocols. In order to work out a follow-up guide, we have retrospectively evaluated the records of 840 stage I cutaneous melanoma patients surgically treated and followed during the postoperative period in the Division of Plastic Surgery of the University of Florence from 1975 to 1992. METHODS: We evaluated the patients' records by analyzing time, pathway and site of any first recurrence in relation to the main prognostic factors such as patient sex, site, histological type and depth of invasion of each primary melanoma. A statistical analysis was performed. RESULTS: To summarize, the salient results were the following: 80% of relapses occurred in the first 3 years and they occurred significantly earlier when the primary melanoma was localized in the trunk and significantly later when the melanoma was localized in the lower limbs and for < 1.5 mm lesions. The first recurrence occurred earlier by the lymphatic than by the hematic pathway regarding the overall number of patients. The hematic pathway was the most frequent (with respect to the overall percentage of hematic metastases) for the melanomas localized in the head and neck region and for lentigo malignant melanomas, whereas the lymphatic pathway was most frequent for melanomas of the lower limbs and > 3 mm in thickness. CONCLUSIONS: We suggest a follow-up schedule taking into consideration the postoperative behavior of stage I cutaneous melanoma patients (in terms of time and pathway of the first recurrence) in relation to the site and depth of invasion of the tumor.

Female↗
PubMed1994

Progesterone 5-alpha-reduction in neuronal and in different types of glial cell cultures: type 1 and 2 astrocytes and oligodendrocytes.

Progesterone, like testosterone, can be converted in the brain into 5-alpha-reduced metabolites (5-alpha-pregnan-3,20-dione, DHP; 5-alpha-pregnan-3-alpha-ol-20-one, THP). Recently we have shown that testosterone is 5-alpha-reduced to DHT mainly in neurons, while glial cells possess this enzymatic activity only in limited amounts. On the other hand, a glial cell type (type 1 astrocytes) is almost exclusively responsible for the further metabolism of DHT into 3-alpha-diol. The aim of the present studies was that of evaluating the formation of the 5-alpha-reduced metabolites of progesterone in cultures of neurons, type 1 and 2 astrocytes and oligodendrocytes. The data here presented indicate that, similarly to what happens when testosterone is used as the substrate, the 5-alpha-reductase which metabolizes progesterone shows a significantly higher activity in neurons than in glial cells; however, also type-1 and type-2 astrocytes as well as oligodendrocytes possess some ability to 5-alpha-reduce progesterone. On the contrary, the 3-alpha-hydroxysteroid dehydrogenase (3-alpha-HSD), the enzyme which converts DHP into THP, appears to be mainly present in type-1 astrocytes; much lower levels of this enzyme are present in neurons and in type-2 astrocytes. At variance with the previous results obtained utilizing androgens as precursors, oligodendrocytes show a considerable 3-alpha-HSD activity, even if this is statistically lower than that present in type-1 astrocytes. The existence of isoforms of the enzymes involved in androgen and progesterone metabolism may explain these data.

3-Hydroxysteroid Dehydrogenases↗
PubMed1994

Estrogen receptor is expressed in different types of glial cells in culture.

Estrogens derived from the aromatization of androgens are believed to be responsible for the induction of the sexual differentiation of the CNS interacting with specific estrogen receptors (ER) present in developing neurons. However, the brain cellular distribution of ER is not so well documented. The aim of this study was to investigate the qualitative and quantitative expression of ER mRNA in well characterized cultures of rat type 1 and type 2 astrocytes and of oligodendrocytes by polymerase chain reaction. A series of amplifications with a set of primers spanning along the entire ER mRNA was utilized in the different types of glial cells, in a positive control (uterus), and in a negative control (SK-N-BE cell line) previously shown to be devoid of ER. The data obtained show that ER mRNA is expressed in all three types of glial cell analyzed in almost equal amounts, which are 25-50 times lower than those in the uterus. The mRNA expressed in the glia is homologous with that expressed in the uterine tissue.

Animals↗
PubMed1994

Proliferative, phenotypic and functional and molecular characteristics of tumour-infiltrating lymphocytes obtained from unselected patients with malignant melanomas and expanded in vitro in the presence of recombinant interleukin-2.

Tumour-infiltrating lymphocytes (TIL) derived from several histotypes, including melanoma, can be expanded in vitro in the presence of recombinant interleukin-2 (rIL-2) and infused as part of experimental adoptive immunotherapy protocols. Several authors have described the isolation and the expansion of TIL, but little is known about the actual proportion of unselected melanoma biopsies that give rise to 'positive' TIL cultures. We evaluated the proliferative, phenotypic, functional and molecular characteristics of cultured TIL obtained from 26 patients with metastatic melanomas, eligible for inclusion in a protocol of adoptive immunotherapy. Sixteen proliferating cultures were obtained. All TIL belonged to the T cell lineage and were characterized by a wide range of cytolytic activity against autologous and, to a lesser extent, allogeneic melanoma cells. Molecular analysis of the constant region of T cell receptor-beta (TCR-beta) showed an oligoclonal population of TIL in the majority of expanded samples, indicating that a selected subset of lymphocytes present in the tumour site could be expanded in vitro. These features, together with the high number of proliferating samples, make these populations of TIL suitable for infusion into patients with advanced disease.

Adult↗
PubMed1994

Steroid binding and metabolism in the luteinizing hormone-releasing hormone-producing neuronal cell line GT1-1.

LHRH synthesis and release are modulated in vivo by gonadal steroids. Although immunocytochemical and autoradiographic studies failed to detect appreciable amounts of estrogen or androgen receptor in LHRH-producing neurons, the recent finding that the promoter region of the LHRH gene contains several steroid hormone-responsive elements indicates a possible direct effect of sex steroids on these specialized neurons. The immortalized LHRH-producing neuronal cell line, GT1, which became recently available, may allow the study of LHRH dynamics. The presence of specific binding sites for estrogen and androgens as well as the presence of the two major enzymatic pathways involved in modulation of androgen action (the 5 alpha-reductase/3 alpha-hydroxysteroid dehydrogenase and the aromatase) have been studied in the GT1-1 clone. High affinity, low capacity binding sites for [3H]estradiol (Kd, 0.11 nM; binding capacity, 6.2 fmol/mg protein) and for a ligand of the androgen receptor, [3H]R1881 (Kd, 0.054 nM; binding capacity, 9.58 fmol/mg protein), have been identified in this cell line. A 2-fold induction of androgen-binding sites has been observed after 3 days of treatment of GT1-1 cells with estradiol (1 microM), indicating that the estradiol binding is probably linked to a functional estrogen receptor. Aromatase and 5 alpha-reductase/3 alpha-hydroxysteroid dehydrogenase activities have been also tested in GT1-1 cells. Under the culture conditions adopted, no detectable aromatization of [1 beta 3H]delta 4-androstenedione to estrone was observed using the tritiated water method. On the other hand, GT1-1 cells efficiently converted testosterone into dihydrotestosterone and subsequently into 5 alpha-androstan-3 alpha,17 beta-diol. In conclusion, GT1-1 cells possess several elements of the machinery through which sex steroids may influence LHRH dynamics.

3-Hydroxysteroid Dehydrogenases↗
PubMed1994

The anterior pituitary gland as a possible site of action of kainic acid.

The purpose of the present study was to analyze the direct effect of kainic acid (KA), an agonist of L-Glutamate, on the secretion of LH and FSH from anterior pituitary (AP) of male rats perifused in vitro. At low concentrations (1 microM), KA was able to stimulate the release of both gonadotropins from AP of 50-day-old male rats, but the response to subsequent stimuli was markedly impaired. This, however, was not due to a neurotoxic action of KA, but seemed rather suggestive of a down-regulation or desensitization of KA receptors. The stimulatory action of KA on LH and FSH secretion was age-dependent, since the agonist was completely ineffective on the AP of 75-day- and 18-month-old male rats. DNQX (6,7-dinitroquinoxaline-2,3-dione), a specific antagonist of the KA receptor subtype, was able to block the KA-induced gonadotropin secretion; similarly, AP-5 (2-amino-5-phosphonovalerate), a competitive NMDA receptor antagonist, prevented the stimulatory effect of KA on LH and FSH release. An interaction between the opiatergic and the excitatory aminoacid (EAA) systems emerged from the observation that pulses of KA applied to AP of 50-day-old male rats during a continuous perifusion with a medium containing morphine (5 microM) failed to increase gonadotropin secretion. These results indicate that KA can, at low concentrations, directly stimulate LH and FSH secretion by acting at AP level; this effect disappears with progression of age, and might be exerted both through NMDA and non-NMDA receptor subtypes. Finally, the results provide evidence that opioids and excitatory aminoacids might influence gonadotropin secretion from AP by acting in opposite directions.

2-Amino-5-phosphonovalerate↗
PubMed1994

Effect of ovariectomy on the binding characteristics of hypothalamic mu opioid receptors in the rat.

The present experiments have been performed to analyze whether ovariectomy, performed in adult female rats, might influence the binding characteristics of hypothalamic mu binding sites. To this purpose, different groups of ovariectomized females have been killed by decapitation 1, 3, 7, 15 and 21 days after surgery, and the binding characteristics (Bmax and Kd) of the mu receptors have been determined in their hypothalami. Dihydromorphine was used as a specific mu ligand. The data obtained have been compared with those derived from the hypothalami of adult female rats in the morning of proestrus, when the density of hypothalamic mu opioid receptors reaches its highest concentration. Blood from trunk vessels was also collected for the evaluation of serum LH levels by radioimmunoassay. The results obtained have shown that one day after ovariectomy, the number of hypothalamic mu opioid binding sites is as high as that found in proestrus animals. The number of these binding sites tends to decrease on days 3, 7 and 15 after surgery. On day 21 after gonadectomy, the number of mu binding sites results to be significantly lower than that observed in the hypothalami of intact female rats in the morning of proestrus and in the hypothalami of female animals castrated since one day. The value of the Kd did not show any variation on days 1, 3, 15 and 21; however, a significant increase of this parameter (which indicates a decrease of the affinity of the ligand for its receptors) was observed on day 7 after ovariectomy.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Interference of thyroperoxidase on immuno-cytochemical determination of steroid receptors in thyroid tissue.

The presence of sexual steroid receptor proteins in thyroid tissue has been previously demonstrated by biochemical means. The aim of this study was to determine the estrogen (ER) and progesterone (PR) receptors in malignant (12 papillary and 1 follicular carcinoma) and nonmalignant (19 multinodular goiters, 1 Graves' disease, 1 Hashimoto's thyroiditis) thyroid diseases using immunocytochemical assay employing monoclonal anti-ER and anti-PR antibodies and the peroxidase-antiperoxidase technique. Positive results were obtained in 24/34 (70%) for ER (ER-ICA+) and 22/34 (64%) for PR (PR-ICA+). To evaluate the possible interference of thyroperoxidase in the immunostaining, in consecutive sections of a positive specimen, primary antibody or primary antibody plus bridging antibody or PAP complex was omitted. Using these modified procedures, staining distribution was similar to that obtained by the standard procedure: in contrast, no staining was found in the positive control, i.e. a breast cancer specimen. The inhibition of the endogenous peroxidase caused a loss of staining in both the standard and modified procedures on thyroid specimens; no staining modification was obtained in the positive control. These results suggest that the staining observed in thyroid tissue is not specific and related to the activity of thyroperoxidase on chromogen solution. The complete loss of staining after peroxidase inhibition appears to be in contrast with the results obtained by biochemical method, and different antigenicity of thyroid receptors in comparison with breast receptors may explain this discrepancy.

Adult↗
PubMed1993

Androgen and progesterone metabolism in the central and peripheral nervous system.

This paper summarizes the most recent data obtained in the authors' laboratory on the metabolism of testosterone and progesterone in neurons, in the glia, and in neuroblastoma cells. The activities of the 5 alpha-reductase (the enzyme that converts testosterone into dihydrotestosterone, DHT), and of the 3 alpha-hydroxysteroid dehydrogenase (the enzyme that converts DHT into 5 alpha-androstane-3 alpha, 17 beta-diol, 3 alpha-diol) have been first evaluated in primary cultures of neurons, oligodendrocytes and type-1 and -2 astrocytes, obtained from the fetal or neonatal rat brain. All the cultures were used on the fifth day. The formation of DHT of 3 alpha-diol was evaluated incubating the different cultures with labeled testosterone or DHT as substrates. The results obtained indicate that the formation of DHT takes place preferentially in neurons; however, type-2 astrocytes and oligodendrocytes also possess considerable 5 alpha-reductase activity, while type-1 astrocytes show a much lower enzymatic concentration. A completely different localization was observed for 3 alpha-hydroxysteroid dehydrogenase; the formation of 3 alpha-diol appears to be prevalently, if not exclusively, present in type-1 astrocytes; 3 alpha-diol is formed in very low yields by neurons, type-2 astrocytes and oligodendrocytes. The compartmentalization of two strictly correlated enzymes (5 alpha-reductase and 3 alpha-hydroxysteroid dehydrogenase) in separate central nervous system (CNS) cell populations suggests the simultaneous participation of neurons and glial cells in the 5 alpha-reductive metabolism of testosterone. Subsequently it has been shown that, similarly to what happens when testosterone is used as the substrate, the 5 alpha-reductase which metabolizes progesterone into 5 alpha-pregnane-3,20-dione (DHP) shows a significantly higher activity in neurons than in glial cells; however, type-1 and -2 astrocytes as well as oligodendrocytes also possess some ability to 5 alpha-reduce progesterone. On the other hand, 3 alpha-hydroxysteroid dehydrogenase, the enzyme which converts DHP into 5 alpha-pregnane-3 alpha-ol-20-one, appears to be present mainly in type-1 astrocytes; much lower levels of this enzyme are present in neurons and in type-2 astrocytes. At variance with the previous results obtained using androgens as precursors, oligodendrocytes show considerable 3 alpha-hydroxysteroid dehydrogenase activity, even if this is statistically lower than that present in type-1 astrocytes. The existence of isoforms of the enzyme involved in androgen and progesterone metabolism is discussed.(ABSTRACT TRUNCATED AT 400 WORDS)

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗
PubMed1993

Distribution of aromatase activity in cultured neurons and glia cells.

Aromatase plays a crucial role in the mechanism of action of testosterone in the central nervous system. Nevertheless, the exact cellular localization of this enzymatic complex within the different cell populations of the brain is still uncertain. In the experiments described here the presence of aromatase (assayed by the tritiated water method) has been evaluated in the two main cellular components of the brain: neurons and glia. Neurons, mixed glial cells, type 1 astrocytes, were obtained in cultures; oligodendrocytes were prepared by gradient ultracentrifugation. The results indicate that, among the different cells tested, only neurons possess a significant degree of aromatase activity, while the enzymatic activity is extremely low in mixed glial cell and in astrocyte preparations. Oligodendrocytes seem to be completely inactive in this respect.

Animals↗
PubMed1993

Testosterone and progesterone metabolism in the human neuroblastoma cell line SH-SY5Y.

The ability of the human neuroblastoma cell line SH-SY5Y to metabolize androgens and progesterone was studied by incubating the cells in the presence of labeled testosterone (T) or progesterone (P) to measure, respectively, the formation of dihydrotestosterone (DHT) or dihydroprogesterone (DHP) (5 alpha-reductase activity). The 3 alpha-hydroxysteroid dehydrogenase activity was studied by evaluating the conversion of labeled DHT into 5 alpha-androstan-3 alpha, 17 beta-diol (3 alpha-diol). The results show that undifferentiated neuroblastoma cells possess a significant 5 alpha-reductase activity, as shown by the considerable conversion of T into DHT; moreover, this enzymatic activity seems to be significantly stimulated following cell differentiation induced by the phorbol ester TPA, but not after differentiation induced by retinoic acid (RA). The 5 alpha-reductase(s) present in SH-SY5Y cells is also able to convert P into DHP. In undifferentiated cells, this conversion was about 8 times higher than that of T into DHT. Under the influences of TPA and RA, the formation of DHP followed the same pattern observed for the formation of DHT. SH-SY5Y cells also appear to possess the enzyme 3 alpha-hydroxysteroid dehydrogenase, since they are able to convert DHT into 3 alpha-diol. This enzymatic activity is not altered following TPA-induced differentiation and appears to be decreased following treatment with RA. It is suggested that the SH-SY5Y cell line may represent a useful "in vitro" model for the study of the mechanisms involved in the control of androgen and P metabolism in nervous cells.

20-alpha-Dihydroprogesterone↗
PubMed1993

Binding characteristics of hypothalamic mu opioid receptors throughout the estrous cycle in the rat.

This study presents a detailed analysis of the binding characteristics of hypothalamic mu opioid receptors during the different phases of the estrous cycle of the female rat. Different groups of adult female rats with a regular 4-day estrous cycle were killed by decapitation at 10.00 h of diestrus days 1 and 2, at 10.00, 12.00, 14.00, 16.00, 18.00 and 20.00 h of the day of proestrus and at 10.00, 12.00, 14.00, 16.00 and 18.00 h of the day of estrus. At sacrifice, the hypothalami of the animals were dissected out, plasma membrane preparations were obtained and the binding characteristics (Bmax, Kd) of the specific mu opioid ligand dihydromorphine (DHM) on mu opioid receptors were evaluated. Blood has been collected from the trunk vessels to monitor with specific radioimmunoassays serum levels of LH, estradiol and progesterone. The data obtained indicate that in the hypothalamus of female rats with a regular 4-day estrous cycle, the binding characteristics of DHM for mu receptors show important variations during the different phases of the estrous cycle. In general, the number of mu opioid receptors is elevated during the morning of diestrus day 2 and during the day of proestrus being maximal at 14.00 h and declining significantly at 18.00 and 20.00 h of the same day. At estrus, the number of mu receptors appears high at 10.00 and 16.00 h and low at 12.00, 14.00 and 18.00 h. All these variations take place without any significant change of the affinity (Kd) of DHM for the mu receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Differential localization of the 5 alpha-reductase and the 3 alpha-hydroxysteroid dehydrogenase in neuronal and glial cultures.

The activities of the 5 alpha-reductase [the enzyme that converts testosterone into dihydrotestosterone (DHT)] and 3 alpha-hydroxysteroid dehydrogenase [the enzyme that converts DHT into 5 alpha-androstan-3 alpha, 17 beta-diol (3 alpha-diol)] have been evaluated in primary cultures of neurons, oligodendrocytes, and type 1 and 2 astrocytes obtained from fetal or neonatal rat brain. All cultures were used on the fifth day. The formation of DHT and 3 alpha-diol was evaluated by incubating the different cultures with [14C]testosterone or [14C]DHT as substrates. The results obtained indicate that the formation of DHT takes place preferentially in neurons; however, type 2 astrocytes and oligodendrocytes also possess considerable 5 alpha-reductase activity, while type 1 astrocytes show a much lower enzymatic activity. A completely different localization was observed for 3 alpha-hydroxysteroid dehydrogenase. The formation of 3 alpha-diol appears to be mainly, if not exclusively, present in type 1 astrocytes. 3 alpha-Diol is formed in very low yields by neurons, type 2 astrocytes and oligodendrocytes. The compartmentalization of two strictly correlated enzymes (5 alpha-reductase and 3 alpha-hydroxysteroid dehydrogenase) in separate central nervous system cell populations suggests the simultaneous participation of neurons and glial cells in the 5 alpha-reductive metabolism of testosterone and possibly other hormonal steroids (e.g. progesterone, corticoids, etc.).

3-Hydroxysteroid Dehydrogenases↗