Myelin changes in the rats CNS following intraventricular injection of serum

Summary Normal human or rat serum administered by intraventricular injection induced demonstrable changes in the rat CNS myelin as seen from an increased recovery of dissociated myelin (DM), i.e. a myelin-related low density membrane fragments, from the tissue homogenates. The yield of DM reached a maximum on the third postinjection day and returned to the control level by day 5. In spite of the increased recovery of DM, no physico-chemical alternations in myelin isolates and no histological abnormalities in the tissue could be detected. The production of DM seems to be a sensitive index of serum-induced alteration of the myelin sheath.Acknowledgments. The authors thank J. Mellach and P. Salinas for their skillful technical assistance. The study was supported by the Danish Multiple Sclerosis Society.     

Myelination in the hippocampus during development and following lesion

Myelin is crucial for the stabilization of axonal projections in the developing and adult mammalian brain. However, myelin components also act as a non-permissive and repellent substrate for outgrowing axons. Therefore, one major factor which accounts for the lack of axonal regeneration in the mature brain is myelin. Here we report on the appearance of mature, fully myelinated axons during hippocampal development and following entorhinal lesion with the myelin-specific marker Black Gold. Although entorhinal axons enter the hippocampal formation at embryonic day 17, light and ultrastructural analysis revealed that mature myelinated fibers in the hippocampus occur in the second postnatal week. During postnatal development, increasing numbers of myelinated fibers appear and the distribution of myelinated fibers at postnatal day 25 was similar to that found in the adult. After entorhinal cortex lesion, a specific anterograde denervation in the hippocampus takes place, accompanied by a long-lasting loss of myelin. Quantitative analysis of myelin and myelin breakdown products at different time points after lesion revealed a temporally close correlation to the degeneration and reorganization pha-ses in the hippocampus. In contrast, electroconvulsive seizures resulted in brief demyelination and a faster recovery time course. In conclusion, we could show that the appearance of mature axons in the hippocampus is temporally regulated during development. In the adult hippocampus, demyelination was found after anterograde degeneration and also following seizures, suggesting that independent types of insult lead to demyelination. Reappearing mature axons were found in the hippocampus following axonal sprouting. Therefore, our quantitative analysis of mature axons and myelination effectively reflects the readjusted axonal density and possible electrophysiological balance following lesion. Received 22 December 2003; received after revision 11 February 2004; accepted 17 February 2004     

Ca2+ uptake and distribution in alloxan-diabetic rat arterial and venous smooth muscle

Summary This report demonstrates that in experimental diabetes mellitus (DM) calcium uptake and its distribution is altered in rat aortic but not in portal venous smooth muscle. Results are interpreted as consequences of increased calcium binding by aortic smooth muscle in experimental DM, which could account for the hyporeactivity of alloxan diabetic rat aorta reported previously.Supported by NIH grant No.HL-18015 and NIDA grant No. DA-02339.     

35S-sulphate incorporation into myelin sulphated mucopolysaccharides during rat brain development.

Rat brain myelin acid mucopolysaccharides (AMPS) incorporate 15%, 8%, 5.5% and 4% of total associated 35S-sulphate, 14, 21, 30 and 75 days after birth, respectively. The course of 35S-sulphate incorporation into total rat brain mucopolysaccharides, as well in those from myelin, had a similar feature with peak on the 2nd week and a significant decrease on the 3rd and 4th week postnatally.     

35S-Sulphate incorporation into myelin sulphated mucopolysaccharides during rat brain development

Summary Rat brain myelin acid mucopolysaccharides (AMPS) incorporate 15%, 8%, 5.5% and 4% of tojtal associated³⁵S-sulphate, 14, 21, 30 and 75 days after birth, respectively. The course of³⁵S-sulphate incorporation into total rat brain mucopolysaccharides, as well in those from myelin, had a similar feature with peak on the 2nd week and a significant decrease on the 3rd and 4th week postnatally.     

Fibronectin in tissue regeneration: timely disassembly of the scaffold is necessary to complete the build

Tissue injury initiates extracellular matrix molecule expression, including fibronectin production by local cells and fibronectin leakage from plasma. To benefit tissue regeneration, fibronectin promotes opsonization of tissue debris, migration, proliferation, and contraction of cells involved in the healing process, as well as angiogenesis. When regeneration proceeds, the fibronectin matrix is fully degraded. However, in a diseased environment, fibronectin clearance is often disturbed, allowing structural variants to persist and contribute to disease progression and failure of regeneration. Here, we discuss first how fibronectin helps tissue regeneration, with a focus on normal cutaneous wound healing as an example of complete tissue recovery. Then, we continue to argue that, although the fibronectin matrix generated following cartilage and central nervous system white matter (myelin) injury initially benefits regeneration, fibronectin clearance is incomplete in chronic wounds (skin), osteoarthritis (cartilage), and multiple sclerosis (myelin). Fibronectin fragments or aggregates persist, which impair tissue regeneration. The similarities in fibronectin-mediated mechanisms of frustrated regeneration indicate that complete fibronectin clearance is a prerequisite for recovery in any tissue. Also, they provide common targets for developing therapeutic strategies in regenerative medicine.     

Myelin architecture: zippering membranes tightly together

Rapid nerve conduction requires the coating of axons by a tightly packed multilayered myelin membrane. In the central nervous system, myelin is formed from cellular processes that extend from oligodendrocytes and wrap in a spiral fashion around an axon, resulting in the close apposition of adjacent myelin membrane bilayers. In this review, we discuss the physical principles underlying the zippering of the plasma membrane of oligodendrocytes at the cytoplasmic and extracellular leaflet. We propose that the interaction of the myelin basic protein with the cytoplasmic leaflet of the myelin bilayer triggers its polymerization into a fibrous network that drives membrane zippering and protein extrusion. In contrast, the adhesion of the extracellular surfaces of myelin requires the down-regulation of repulsive components of the glycocalyx, in order to uncover weak and unspecific attractive forces that bring the extracellular surfaces into close contact. Unveiling the mechanisms of myelin membrane assembly at the cytoplasmic and extracelluar sites may help to understand how the myelin bilayers are disrupted and destabilized in the different demyelinating diseases.     

Ultrastructural investigation of fetal rat brain hemisphere tissue in nonadherent stationary organ culture

Summary Fetal rat brain fragments grown in nonadherent stationary organ culture for 50 days have been investigated ultrastructurally. Synaptogenesis and myelin formation occurred at the same time as the corresponding time-dependent events in the developing brain in vivo. Intermediate junctions were observed between cellular processes lining a central cavity in the fragments and later associated with astrocytes at the surface. Gap junctions and tight junctions were also present. In some fragments cilia were observed in the central cavity. Subependymal basement membrane labyrinths were observed in all fragments after 10 days in culture. The ultrastructural characteristics and the tissue-like structure in general were preserved for at least 50 days in this tissue culture system. The brain fragments may therefore be a valuable supplement to existing culture methods for nervous tissue.Research fellow from the Norwegian Research Council for Sciences and Humanities. This work was supported by the Norwegian Cancer Society.     

Myelin sheaths: glycoproteins involved in their formation,maintenance and degeneration

Myelin sheaths are formed around axons by extending, biochemically modifying and spiraling plasma membranes of Schwann cells in the peripheral nervous system (PNS) and oligodendrocytes in the central nervous system (CNS). Because glycoproteins are prominent components of plasma membranes, it is not surprising that they have important roles in the formation, maintenance and degeneration of myelin sheaths. The emphasis in this review is on four integral membrane glycoproteins. Two of them, protein zero (P0) and peripheral myelin protein-22 (PMP-22), are components of compact PNS myelin. The other two are preferentially localized in membranes of sheaths that are distinct from compact myelin. One is the myelin-associated glycoprotein, which is localized at the inside of sheaths where it functions in glia-axon interactions in both the PNS and CNS. The other is the myelin-oligodendrocyte glycoprotein, which is preferentially localized on the outside of CNS myelin sheaths and appears to be an important target antigen in autoimmune demyelinating diseases such as multiple sclerosis. Received 8 April 2002; received after revision 13 May 2002; accepted 22 May 2002     

Development and hormonal control of thioredoxin and the thioredoxin-reductase system in the rat liver during the perinatal period

The development and hormonal regulation of thioredoxin and of the thioredoxin-reductase system were investigated during the perinatal period in rat liver. An immunological procedure was developed in order to quantify thioredoxin in fetal and neonatal hepatocytes. Both immunoreactive thioredoxin and thioredoxin-reductase activity appeared on day 16.5 of pregnancy. The level of immunoreactive thioredoxin increased during the late fetal period, and its level was the same 24 h after birth. Moreover, its development was not subjected to hormonal regulation by corticosteroids and glucagon. In contrast, thioredoxin-reductase activity increased 3 times during the late fetal period and presented a marked increase 24 h after birth. In the absence of glucocorticoids there was no increase in the level of thioredoxin reductase, while administration of hydrocortisone acetate and glucagon to fetuses prematurely evoked its activity. This study suggests that if thioredoxin acts physiologically, this activity is related to the state of reduction of the molecule rather than to the total concentration in the liver.     

Development and hormonal control of thioredoxin and the thioredoxin-reductase system in the rat liver during the perinatal period

Summary The development and hormonal regulation of thioredoxin and of the thioredoxin-reductase system were investigated during the perinatal period in rat liver. An immunological procedure was developed in order to quantify thioredoxin in fetal and neonatal hepatocytes. Both immunoreactive thioredoxin and thioredoxin-reductase activity appeared on day 16.5 of pregnancy. The level of immunoreactive thioredoxin increased during the late fetal period, and its level was the same 24 h after birth. Moreover, its development was not subjected to hormonal regulation by corticosteroids and glucagon. In contrast, thioredoxin-reductase activity increased 3 times during the late fetal period and presented a marked increase 24 h after birth. In the absence of glucocorticoids there was no increase in the level of thioredoxin reductase, while administration of hydrocortisone acetate and glucagon to fetuses prematurely evoked its activity. This study suggests that if thioredoxin acts physiologically, this activity is related to the state of reduction of the molecule rather than to the total concentration in the liver.     

成年大鼠脊髓损伤后内源性神经前体细胞的增殖与分化规律的研究

目的观察成年大鼠脊髓损伤后内源性神经前体细胞的增殖与分化,探讨内源性神经前体细胞的自然变化规律。方法制作脊髓压迫损伤模型,Brdu腹腔注射标记神经前体细胞,免疫荧光法(Immunofluoreseence)检测大鼠脊髓Brdu、GFAP、MBP阳性细胞数的变化。结果 1)正常组可观察到少量Brdu阳性细胞,脊髓损伤后Brdu阳性细胞显著增加(p0.05),并在第7天达到最大值,21天时仍高水平表达。2)正常组可见少量Brdu/GFAP和Brdu/MBP阳性细胞,脊髓损伤后Brdu/GFAP,Brdu/MBP双标阳性细胞数显著增加(p0.05)。结论脊髓损伤后神经前体细胞的数量在第7天达到最大值,我们认为,一周内可能是神经前体细胞增殖分化调控的关键时期。此外,新生星形胶质细胞和少突胶质细胞大量增殖,并与神经前体细胞的迁移、后肢功能恢复表现出一定的同步性,提示新生胶质细胞可能参与了脊髓损伤后神经功能的修复作用。     

外源性干细胞因子对糖尿病胃轻瘫大鼠胃窦Cajal细胞的影响

目的探讨外源性干细胞因子（SCF）能否改善糖尿病胃轻瘫（DGP）大鼠胃窦Cajal细胞（ICC）的异常病变。方法sD大鼠随机分为实验组和对照组，实验组腹腔注射链脲佐菌素（STZ）50mg／kg并高糖高脂饲料不规律喂养构建DGP模型，对照组腹腔注射等量生理盐水并普通饲料规律喂养。成模后随机分为糖尿病组（DM组）、糖尿病＋外源性干细胞因子治疗组（DM＋SCF组）；DM＋SCF纽腹腔注射SCF0．4ug／（kg·d），共15天，对照组和DM纽每天腹腔注射等量的磷酸盐缓冲液（pH=7．4）。RT-PCR、Wester Dblot检测胃窦组织中SCF及c—kit的mRNA及蛋白表达；免疫组化、透射电镜观察胃窦组织中Cajal细胞的变化。结果DM组大鼠胃窦组织SCF及c—kit的mRNA及蛋白表达下降，Cajal细胞数量减少，细胞内线粒体肿胀、内质网扩张；予外源性SCF干预后，胃窦组织中SCF及C—kjf的mRNA及蛋白表达上调，caial细胞数量增多、超微结构明显改善。结论外源性SCF能在一定程度上改善或逆转糖尿病胃轻瘫大鼠胃窦Cajal细胞的异常病变：     

早期营养不良对SD大鼠海马AchE和ChAT表达的影响

目的探讨生命早期营养不良对21天雄性sD大鼠中枢AchE和ChAT的影响。方法从妊娠晚期到哺乳期末给予母鼠半量饲料,造成子鼠营养不良,取哺乳期末雄性子鼠脑海马组织进行测量。结果免疫组化结果示：实验组海马CA3区ChAT光密度低于对照组（P〈0．05）,在CA!区无差异（P〉0．05）;而两纽间CAl和CA3区AchE光密度均无差异（P〉0．05）;RT—PCR示：实验组海马ChAT、AchEmRNA少于对照组（P〈0．05）。结论早期营养不良能导致子鼠海马及中枢胆碱能系统发育障碍,可能会影响成年后大鼠学习记忆能力。     

5-Hydroxytryptamine binding to butanol extracts from myelin fragments

Summary The myelin fraction of rat brain stem was treated with butanol-water mixtures, and the extracted proteolipids were separated by Sephadex LH₂₀ column chromatography. 2 peaks of proteolipids eluted in chloroform-methanol 4/1 showed the binding capacity for C¹⁴·5-HT. This finding suggests the necessity of the more careful investigations for the probability of proteolipids as receptor proteins in the central nervous system.     

Pulmonary di-and-triacylglycerols during the perinatal development of the rat

Summary Diacylglycerol (DG) and triacylglycerol (TG) levels in rat lung tissue were determined from day 17 of gestation to day 10 post partum and studied in parallel with ultrastructural differentiation. The DG level, although rather low at all measured stages, rose significantly between days 17 and 19 and at birth. TG level increased steadily during the whole studied period and especially between days 17 and 19 and at birth. In DG as well as in TG, saturated fatty acids were predominant. The rising of TG levels paralleled the appearance and accumulation of lipid vacuoles in mesodermal cells lying in contact with type II cells. The possible role of these cells is discussed.     

The effect of bovine myelin basic protein on uptake and release of H3-labelled 5-hydroxytryptamine,L-noradrenalin and γ-aminobutyric acid in rat cortex slices

Summary At concentrations above 10^–5 M myelin basic protein (MBP) induced a small inhibition of the uptake of H³-5HT and H³-NA into rat cortex slices. Release of 5HT, NA and Gaba was not affected by 10^–5 M MBP.Acknowledgment. The authors wish to thank W. Bucher for the preparation of MBP.     

Synergism between long-acting bromocryptine microcapsules and cyclosporine A in the prevention of various autoimmune diseases in rats

Pre-treatment of male Sprague-Dawley rats with long-acting bromocryptine microcapsules (CBLA) significantly inhibited the arthritic response to Freund's complete adjuvant and reduced weight loss, thymolysis, splenomegaly and leukocytosis. In the prevention of adjuvant arthritis (AA), the combination of CBLA plus sub-optimal doses of cyclosporine A (CsA) was more efficient than either of the drugs alone. Sub-optimal doses of CsA were 0.1 and 1.0 mg/kg/day s.c. for 5 days. Furthermore, CBLA alone did not decrease the incidence of experimental allergic uveitis (EAU) in the male Lewis rats. Low-dose CsA reduced the incidence of uveitis by 50%, and with the addition of CBLA, 100% of rats were protected. Low-dose CsA was 2 mg/kg/day i.m. for 14 days. Long-term treatment of male Sprague-Dawley rats with CBLA alone reduced the incidence and severity of spontaneous autoimmune periateritis nodosa (PN) in a dose-dependent manner; CsA was less potent than CBLA, and only additive effects were obtained. Finally, for the prevention of spontaneous autoimmune insulin-dependent diabetes (DM), the administration of CBLA did not improve the effect of a low-dose CsA in male BB rats. Nevertheless, a delay in onset of DM could be achieved. A sequential therapy using CsA plus CBLA clearly showed beneficial effects. The dose of CsA was 10 mg/kg p.o. 6 days/week for 21 weeks. Compared with Sprague-Dawley or Lewis male rats, BB male rats showed only weak prolactin suppression after the same doses of CBLA. It is suggested that the use of CBLA may be particularly beneficial in autoimmune disorders. The effectiveness of the combination therapy CBLA plus CsA, however, was dependent on the model considered. Various factors could play a role: (1) the different ways of administering CsA (s.c. in AA, i.m. in EAU and PN, oral in DM); (2) strain-dependency in the capacity of CBLA to suppress Prl secretion; and(3) at least in the BB rats, the transient increase of CsA bioavailibility which was possibly induced by CBLA.     

Ontogenetic analysis of the theta rhythm during paradoxical sleep in rats]

In the rat, an ontogenetic study of theta rhythm during paradoxical sleep shows a clear evolution between the 14th and the 30th postnatal day. During this period, its mean frequency increases from 5 to 7 cycles per second, and its spectrum shows significant changes. These parameters (mean frequency and spectral components) of the theta rhythm could be a useful index of brain maturation in the rat.     

Vagal afferent innervation in regenerated rat liver

The possible presence of neural sprouting in the afferent neurons of regenerated rat liver after hepatectomy was investigated by retrograde transport of horseradish peroxidase. This experiment was carried out to see if the increase in hepatic parenchyma could provide an adequate stimulus for the sprouting process. The study was limited to the vagal afferents, particularly the left ones, because they are the principal contributors to hepatic afferent innervation in the rat. The results show that neural sprouting does not occur in regenerated rat liver after 3 weeks. In fact, the number of intensely labeled neurons in the left nodose ganglia of hepatectomized rats was significantly smaller than in controls. This could be due to a lessened availability of horseradish peroxidase to nerve terminals because of the increased non-innervated hepatic mass. There was no difference between right nodose ganglia neurons in hepatectomized and control animals. This could be a consequence of their possible distribution in hepatic areas not involved in the regenerative process.