[Significance of CRF and dopamine receptors in amygdala for reinforcing effects of opiates and opioids on self-stimulation of lateral hypothalamus in rats].

Bipolar electrodes were implanted in the lateral hypothalamus in a group of 44 Wistar male rats in order to study self-stimulation reaction in the Skinner box. Simultaneously, microcanules were implanted into the central nucleus of the amygdala to inject the drugs (1 microl per injection). The blockade of corticoliberin (CRF) receptors (astressin, 1 microg) or Na+influx currents (xycaine or lidocain 1 microg) by the intrastructural administration of drugs into the amygdala decreased self-stimulation reaction of the lateral hypothalamus in rats by 29-55%. The inhibition of D1 and D2 dopamine receptors in the amygdala with SCH23390 (1 microg) or sulpiride (1 microg) respectively, also reduced self-stimulation but to a lower degree. On the background of blockade of CRF (astressin) and dopamine (sulpiride) receptors as well as sodium influx ionic currents (lidocain) in the amygdala neurons, psychomotor stimulant amphetamine (1 mg/kg) and barbiturate sodium ethaminal (5 mg/kg) retained their psychoactivating effect on self-stimulation (+30-37%), while fentanyl (0.1 mg/kg) and leu-enkephaline (0.1 mg/kg) did not produce this effect. Fentanyl moderately activated self-stimulation only after the blockade of D1 dopamine receptors with SCH23390. After the blockade of CRF receptors, leu-enkephaline strengthened its depressant effect on self-stimulation reaction (-89%). Therefore, if the modulating action of amygdala on the hypothalamus is eliminated, the enhancing effects of opiates (fentanyl) and opioids (leu-encephaline) are blocked, but the effects of psychomotor stimulant amphetamine and barbiturate sodium ethaminal are retained.

[Delayed behavioral and morphological consequences in activation of the stress-antistress system in early ontogeny stages in rats].

Wistar rat pups aged 4-10-17 days were injected intraperitoneally with corticoliberin (corticotrophin-releasing hormone, CRH) in doses of 0.5 - 1.0 - 2.0 mg/rat, respectively (single administration for each rat), which activates the stress system, or with 70-kDa heat-shock protein (HSP-70) in doses of 5 - 10 - 20 mg/rat, respectively, which plays the role of intracellular shaperons and possesses antistress properties. The effect of drugs on the emotional and motor behavior was assessed in 5 tests (open field, elevated plus-maze, intruder-resident, Porsolt's depression test, and rotation test) in adult rats 90- 100-days-old. The activation of stress or antistress systems by CRH or HSP-70, respectively, changed the behavior of adult rats. These effects depended on the animal gender, being different in males and females: male rats were more sensitive in Porsolt's depression test, elevated plus-maze anxiety test, and rotation test, while being less sensitive in the open field and intruder-resident tests. These results indicate that the initial sensitivity of males and females with respect to CRH and HSP-70 is also different. The experimental data exhibited correlation with the results of morphological investigation of the limbic structures of the brain. In particular, CRH increased the relief (volume) of neurons of substantia nigra and ventral tegmental region without changing their density, while HSP-70 produced moderate degeneration of neurons and decreased their density. It is suggested the obtained data have to be taken into account in planning and conducting experimental investigations devoted to the influence of various pharmacological agents on the behavior.

[Pharmacodynamic metoclopramide (cerucal) test in differential diagnosis of hypoprolactinemic hypogonadism syndrome in women]. / Farmakodinamicheskaia proba s metoklopramidom (tserukalom) v differentsial'noi diagnostike sindroma giperprolaktinemicheskogo gipogonadizma u zhenshchin.

Roentgenology revealed pituitary macroadenoma in 13 of 67 women with hyperprolactinemia. Craniographic structural abnormalities of the sella turcica suggested pituitary microadenoma in 20 women. Metoclopramide (cerucal) test was performed in the patients and a control group of 9 healthy women. Blood concentrations of prolactin (prolactin, somatotropic thyroid-stimulating hormone) were determined with a radioimmune assay. Significant disorders of the hypothalamo-pituitary control were detected, presenting as depressed and delayed prolactin and TSH response in this assay. The neuroendocrine disorders were most severe in patients with macroprolactinoma who showed a refractory prolactin secretory response to dopaminergic metoclopramide inhibition. The patients with microprolactinomas had a low prolactin response (increments less than 300-400%) and delayed TSH response. The metoclopramide test is recommended for differential diagnosis of hyperprolactinemic syndrome.

[Immunomorphological research on the formation of the extracellular matrix in epithelial cell cultures]. / Immunomorfologicheskoe issledovanie formirovaniia vnekletochnogo matriksa v kul'turakh épitelial'nykh kletok.

Formation of extracellular matrix structures in cultures of rat liver epithelial nontransformed cell line IAR2 was studied with antisera to fibronectin, laminin and type IV collagen by immunofluorescence and immunoelectron microscopy of platinum replicas. Fibronectin formed peripheral spots of variable size some of which outlined free cell edges, as well as fibrils located towards the center of single cells or of cellular islands. Similarly distributed structures were seen in isolated matrices. Codistribution of fibronectin and actin was observed only for the peripheral line of fibronectin spots and marginal circular actin bundle. Basement membrane components. laminin and type IV collagen, formed mainly spots of variable size predominantly beneath the cell or each cell in an island. Occasional fibrils were seen also. Essentially the same results were obtained by immunofluorescence and immunogold electron microscopy. Cytochalasin D treated cells displayed spots of both fibronectin and laminin. The relevance of previously postulated receptor-mediated assembly of extracellular matrix structures to the epithelial cells is discussed.

[Hyperprolactinemia syndrome in men]. / Sindrom giperprolaktinemii u muzhchin.

A total of 122 male subjects suffering from secretory infertility were examined. In 25 of them (20 percent) blood prolactin levels were above the norm. Measurements of basal prolactin levels and of its secretion in metoclopramide test helped distinguish two types of hyperprolactinemia syndrome, differing in the pattern of spermatogenesis disorders. Possible pathogenesis of individual types of hyperprolactinemia in men is discussed. Therapy with dopamine agonists (lisenyl, parlodel) was found most effective in Type I hyperprolactinemia syndrome, associated with essential prolactin hypersecretion and oligospermia.

[Monoclonal antibodies to the proteins of intermediate filaments and of basement membranes in the differential diagnosis of certain forms of human breast tumors]. / Monoklonal'nye antitela k belkam promezhutochnykh filamentov i bazal'nykh membran v differentsial'noi diagnostike nekotorykh form opukholei molochnykh zhelez cheloveka.

Monoclonal antibodies to keratins No. 8 and 17 specific for lining epithelium and myoepithelium of the mammary gland, respectively, as well as to basement membrane laminin, entactin, collagen type IV and heparan sulfate proteoglycan were used to the immunohistochemical analysis of 77 benign and malignant human breast lesions and that of 38 cases in which an intraoperative biopsy diagnosis was difficult. Morphologically similar benign and malignant proliferations were distinguished by keratin expression. In benign lesions both keratins were present, while in malignant ones only keratin No. 8 was expressed. Basement membranes associated with a myoepithelial layer were intact in benign lesions and in situ structures, but they were absent around the vast majority of invasive tumor foci. Basement membrane loss was important in differential diagnosis of benign sclerosing adenosis and cystadenopapilloma from invasive tubular and papillary carcinoma, respectively. Diagnosis of microinvasion in ductal and lobular carcinoma was much easier when combination of antibodies to keratins and basement membrane proteins was used.

[Dopaminergic regulation of the hypothalamo-hypophyseal system in patients with hypophyseal adenomas]. / Dofaminergicheskaia reguliatsiia gipotalamo-gipofizarnoi sistemy u bol'nykh s adenomami gipofiza.

A study was made of the state of the hypothalamohypophyseal system in patients with different types of chromophobe adenoma. Altogether 62 patients were examined using metoclopramide and Parlodel tests (38 patients were with somatotropinoma, 13--with prolactinoma, 11--with somatotropinoma with hyperprolactinemia). The time course of STH, PRL and TSH secretion in the blood was investigated. Dopaminergic blockade in the Metoclopramide test was shown to cause no change in the time course of PRL secretion in patients with prolactinomas, and a PRL reaction in patients with somatotropinomas and adenomas with PRL and STH hypersecretion was sharply decreased. A paradoxical STH reaction (a decrease) in somatotropinomas and considerable disorders in the type of PRL and TSH secretion in all forms of adenomas was defined in the Parlodel test (a dopaminergic agonist). The results obtained suggest considerable disorders in the dopaminergic regulation of the hypothalamohypophyseal system of patients with chromophobe adenomas and the efficacy of the metoclopramide and Parlodel tests in the differential diagnosis of these conditions.

[Process formation and dynamics of the spreading of cultured cells on an extracellular matrix of fibroblasts]. / Formirovanie otrostkov i dinamika rasplastyvaniia kul'tiviruemykh kletok na vnekletochnom matrikse fibroblastov.

Flattening of the normal mouse and human fibroblasts and of the epithelial cells (lines from rat liver IAR-2, trachea of calf foetus FBT and mouse kidney MPTR) was studied on isolated extracellular matrix (EM) formed by human fibroblasts in culture has been studied. EM consisted of fibers, usually parallel to each other. Flattening of the fibroblasts on EM was slower and less even than on glass. Separate processes formed in place of a ring lamella and those processes gradually stretched which were parallel to the EM fibers. Within 24 h fibroblasts were stretched and oriented along the EM fibers. At the EM-glass boundary fibroblasts migrated from EM to glass. Epithelial cells also flattened on EM more slowly and unequally on EM than on glass. Within 24 h, the IAR-2 and FBT cells were less flattened than on glass, acquired a disc-like shape with uneven contours and, sometimes, an oval shape. MPTR cells and their colonies were oriented and stretched along the EM fibers.

[Heterogeneity of basal membranes in human tissues detected by monoclonal antibodies to laminin and entactin]. / Geterogennost' bazal'nykh membran v tkaniakh cheloveka, vyiavliaemaia s pomoshch'iu monoklonal'nykh antitel k lamininu i éntaktinu.

The distribution of basal membrane glycoproteins, laminin and entactin, was studied immunohistochemically by peroxidase-antiperoxidase technique in different adult human organs: kidneys, liver, heart, skin, spleen and ileum. Monoclonal antibody against entactin (ELM2) reacted with all basal membranes. Monoclonal antibody against laminin (LT3.1), however, did not react with basal membranes of arterial smooth muscle cells, or with endothelial basal membranes of renal and hepatic sinusoid endothelium. Thus, LT3.1 antibody has revealed basal membrane heterogeneity by laminin. The possible immunochemical heterogeneity of basal membranes by entactin is also discussed.

[Isolation and characteristics of the extracellular matrix of cultured cells]. / Vydelenie i kharakteristika vnekletochnogo matriksa kul'tiviruemykh kletok.

Sodium deoxycholate extraction was used to isolate extracellular matrix from various cultured cells: human and murine embryonic fibroblasts, epithelial lines of mouse (MPTR), rat (IAR 2 and IAR 20), pig (SPEV) and cow (FBT). Protein composition of the matrix was studied by sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunofluorescence. The matrix morphology was investigated by scanning electron microscopy. In cell lines FBT and MPTR the major component of the matrix was laminin, whereas in other lines and fibroblasts it was fibronectin. The matrix of the majority of lines had a fibrillar structure, and the fibrils usually formed networks. MPTR cells had a punctate matrix composed of laminin and collagen type IV, densely covering the substratum. The treatment of the matrix by hyaluronidase and/or DNAase I did not influence its protein composition. The isolated matrix of different structure and composition may serve a biological substratum in studies of normal tumor cell behavior in tissue culture.

[Fibronectin in cultured mouse sarcoma cells induced by foreign bodies]. / Fibronektin v kul'tiviruemykh kletkakh myshinykh sarkom, indutsirovannykh inorodnymi telami.

Indirect fluorescence was used to study distribution of fibronectin in cultured cells of 5 mouse foreign body-induced sarcomas (PS-4, PS-84, PS-100, PS-103, PS-130) and cells of early (2-3 months postimplantation) capsules surrounding implanted plastic films. Accumulation of pericellular or matrix fibronectin in dense cultures compared to sparse ones was noted in all the lines except PS-84. In confluent cultures of these lines, a three-dimensional fibrillar network of fibronectin typical for fibroblast-like cells was observed. The pattern was very similar to that found in dense cultures of non-neoplastic early capsule cells. The data obtained show that loss of pericellular fibronectin would not serve as a marker of neoplastic transformation in this cellular system.

[Distribution of surface fibronectin in low- and high-density cultures of normal and transformed mouse fibroblasts]. / Raspredelenie poverkhnostnogo fibronektina v redkikh i gustykh kul'turakh normal'nykh i transformirovannykh fibroblastov myshi.

Indirect immunofluorescence was used to study the distribution of surface fibronectin in normal embryonic and in four lines of transformed mouse fibroblasts. Accumulation of the fibronectin matrix with increasing culture density was discovered not only in normal but also in transformed cells. In three out of the four transformed lines studied, fibronectin was found in a considerable amount, which is in agreement with the data obtained with lactoperoxidase iodination. In these neoplastic lines, fibronectin was organized in the form of a fibrillar network like in normal cells. The data provided indicate that the changes in fibronectin content upon neoplastic transformation should be evaluated in dense cultures with maximum expression of this glycoprotein.

[Distribution of fibronectin on the surface of single fibroblasts. The possible mechanism of the assembly of fibronectin structures]. / Raspredelenie fibronektina na poverkhnosti odinochnykh fibroblastov. Vozmozhnyi mekhanizm sborki fibronektinovykh struktur.

Distribution of fibronectin on the surface of lamelloplasm and endoplasm of single cultured mouse embryo fibroblasts was studied by means of indirect immunofluorescence technique. Large fibronectin fibrillae were located on the surface of endoplasm mainly, while "spots" and "streaks" of fibronectin dominated on the surface of lamelloplasm. Colcemid-treated cells had approximately the same distribution whereas cytochalasin B-treated cells were characterized by a spotty fluorescence alone. In contrast, in dense cultures of colcemid-treated cells fibronectin network underwent tearing, while it remained unchanged in cytochalasin B-treated cells. The centripetal movement of the fibronectin--receptor complexes along the cell surface from lamelloplasm into endoplasm (or the natural capping of fibronectin receptors) seems to play an important role in the assembling of fibronectin fibrillae.

[Changes in the surface morphology of normal and transformed mouse fibroblasts following disruption of cell-substrate adhesion]. / Izmeneniia morfologii poverkhnosti normal'nykh i transformirovannykh fibroblastov myshi pri narushenii adgezii kletok k podlozhke.

By means of scanning electron microscopy surface morphology of cultured normal mouse embryo fibroblasts (MEF) and transformed mouse fibroblasts of L strain was studied in the course of alteration of cell-substrate adhesion with proteases, EDTA and urea. The morphology of cell rounding induced by the above agents in MEF and L cells was almost independent on the type of the agent. The rounding of MEF proceeded through three stages and was accompanied by substantial changes of cell surface relief. L cells lacked the intermediate stage (formation of thick processes) during their rounding which proceeded without any changes of cell surface relief. It is suggested that the observed differences are related to the poorer development of the lamelloplasm and microfilaments bundles in the transformed cells ascompared to the normal ones.