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144 Functional and structural analysis of DSD-1-pg glycosaminoglycans
142
133
MORPHOLOGICAL BASIS OF DELTA-SLEEP INDUCING PEPTIDE ANTISTRESSORY ACTION: ULTRASTRUCTURAL CHANGES OF AXOSPINOUS SYNAPSES
ADAPTATION TO HYPOTHERMIA IN AND NON-HIBERNATING HIBERNATING RODENTS: THE REALIZATION OF TWO DIFFERENT STRATEGm
P.Povilaititc,
A.Mendzheritsky, A.Matsionis, G.Kuraev*, I. Pavlov, P.Povilaitite Rostov Regional Bureau of Pathology, Department of Experimental Pathornorphology, *Rostov State University, 344085, Rostov-on-Don, Russia.
A.Ma&ianis,
I.Pavlov,
G.Kuraev*,
A.Mendzhcritsky
Rostov Regional Bureau of Pathology, Dept. of Experimental Pathamorphology, *Rostov State Univ., 344085, Rostov-on-Don, Russia. It has been known that adaptation reactIons of CNS are accompanied by correspondent changes in its ultrastructure. In the present study the morphometric analysis of axospinous synapses of III-V layers of sensorimotor cortex in rats under hypoxia, hyperoxia and under these impacts with preliminary delta-sleep inducing peptide (DSIP) injection is performed. Our measurements indicate that DSIP alone practically does not influence ultrastructure of axospinons synapses, although the decrease of spine area was observed. Both hypoxia and hyperoxia lead to considerable changes of such quantitative parameters of synapses as area of presynaptic bouton, length of active zone, spine area, number of synaptic vesicles, area of mitochondrial profiles, area of mitochondrial profiles/area of axonal ending ratio, form factor of axonal endings, spines and mitochondrial profiles. DSIP injection 1 hour before stress conditions result in appearance of flattened axonal endings stretched out on the spine head. Usually such contacts have two active zones set at angle to each other. The mcrease in synaptic vesicles number and total length of active zone suggests DSIPinduced stimulation of release apparatus work coupled with activation of additional post-synaptic receptors.
Previously we have reported that neocortex of hibematmg animals have highly developed protein synthetic stmctures and complexes of axe-axosornatic and axo-axodendritic contacts - so called “serial synapses”. It is likely that these peculiarities play an important role in adaptation process to low temperature. In ttns connection the present work aims to provide a compar~on studies of sensorimotor cortex of ground squirrels and rats duting normothetia and hypothermia (animals were cooled down to 25°C and were kept at this temperature during hvo hours). The lowering of temperature in rats entails disorganization of cell structures to a variable degree. Many Irregularly shaped lipofuscin granules and secondary lysosomes can be seen in neurons. The majority of dendritic spines are flattened. The reaction of ground squirrels neocorlex to hypothermia, contrastingly, is manifested in activation of receptor medIated exocytosis, lengthening of dendritic spine neck and activation of axe-axonal contacts. The results of our investigation suggests that hibernating and non-hibernating rodents use fundamentally different adaptation strategies while adjusting to low temperatures.
144
136
FUNCTIONAL AND STRUCTURAL ANALYSIS OF DSD-I-PG GLYCOSAMINOGLYCANS
MEMBERS
IDENTIFICATION
A. Clernenr, S. Nadanaka’, K. Masayama’. C. Ueoka*, K. Sugahara*. C. Mandl. and A. Falssner Dcparlmcnl for Neurobiology, University of Hcldclbcrg, 69120 Hcidclberg, FRG; *Department of Blochcmlstry, Kobc Pharmaceutical University, 658 Kobc, Japan
OF
REGC%-1 AND
OF A FAMILY
OF CELL
REGGIE-2,
SURFACE
TWO
PROTEINS
INFXSHANDMAMMALS.
S. Lommel, T. Schulte, M. Jung and C.A.O. Shmmer. Faculty of Biology, University of Konstanz, D-18434 Konstanz, Germany The moncclonal antibody M 802 recognizes a protein on the surface of ne&om retinal ganglion cells (RGCs) and RGC axon.5 tn the continuously growing retina of adult goldfish, which we named reppie. Reggie is re-expressed on all RGCs and RGC axons atIer optic nerve lesion and during axon regeneration suggesting a function of reggie for fiber tract repair. Two cDNAs, reggie-I and reggie-2, were identified in fish, and the lesioninduced upregulation of retie-1 and reggie-2 mRN4.s in RGCs was confirmed by in sifts hybridization. Moreover, we isolated partial rat cDNA clones for reggie-1 and reggie-2 by PCR based
The chondrmtin sulfalc protcoglycan DSD-I-PG has been identificd with tbc monoclonal antibody (mAb) 473HD. The mAb spccifitally rcac& with a structure contained in chondroitin sulfate C (CS C) which is degraded by chondroitinascs ABC and AU. but not by chondroitinase ACII. Thcsc results suppon Ihc mtcrprctation that the cpitope is composed of a dermatan (DS)/CS C hybrid (designated DSD-1 for dcmatan Sulfate dependent). DSD-I-PG IS cxprcssd by ghal precursor cells m a developmentally regulated fashion and Westcm blot analysis of mouse brain extracLr documcntcd maximal expression during early postnatal stages. When coated as substrate. DSD-I-ffi promotes neurite outgrowth of embryonic hippocampal neurons in virro. This suppoaive effccl is pamally blocked by mAb 473HD. which suggests functional rclevancc of the DSD-I glycosaminoglycan (Gag). Further structural analysis indicated that the 473HD-cpitopc requires the sulfation of the Gag. Thus, pharmacological agents. which block sulfation, abolished the epitopc in the DSD-l-PG expressing oligodcndroglial Oli-ncu precursor cell hne, as shown by immunoprecipitation and immunocytochemistry. This conclusion is consistent with competition ELlSAs using various CS of similar srructure, but distinct sulfarc composmon. In order 10 obtam more detailed information about the DSD-1 structure, CS C was digested with chondroitinase ACII. The resulting fragments were SL%Cfractionated by HPLC and tested by competition ELISA. Prchmmary data indicate that the epitopc is contained m a 16mcr fraction and two fractions of higher oligosaccharidcs. Supported by DFG (Fa 159/S-1.2.3)
cloning and obtained evidence for reggie expression in the rat CNS by in siti-hybridization. The characterization and spatiotemporal expression of reggie will be presented. Supported by DFG and BM!3F
xi
133
MORPHOLOGICAL BASIS OF DELTA-SLEEP INDUCING PEPTIDE ANTISTRESSORY ACTION: ULTRASTRUCTURAL CHANGES OF AXOSPINOUS SYNAPSES
ADAPTATION TO HYPOTHERMIA IN AND NON-HIBERNATING HIBERNATING RODENTS: THE REALIZATION OF TWO DIFFERENT STRATEGm
P.Povilaititc,
A.Mendzheritsky, A.Matsionis, G.Kuraev*, I. Pavlov, P.Povilaitite Rostov Regional Bureau of Pathology, Department of Experimental Pathornorphology, *Rostov State University, 344085, Rostov-on-Don, Russia.
A.Ma&ianis,
I.Pavlov,
G.Kuraev*,
A.Mendzhcritsky
Rostov Regional Bureau of Pathology, Dept. of Experimental Pathamorphology, *Rostov State Univ., 344085, Rostov-on-Don, Russia. It has been known that adaptation reactIons of CNS are accompanied by correspondent changes in its ultrastructure. In the present study the morphometric analysis of axospinous synapses of III-V layers of sensorimotor cortex in rats under hypoxia, hyperoxia and under these impacts with preliminary delta-sleep inducing peptide (DSIP) injection is performed. Our measurements indicate that DSIP alone practically does not influence ultrastructure of axospinons synapses, although the decrease of spine area was observed. Both hypoxia and hyperoxia lead to considerable changes of such quantitative parameters of synapses as area of presynaptic bouton, length of active zone, spine area, number of synaptic vesicles, area of mitochondrial profiles, area of mitochondrial profiles/area of axonal ending ratio, form factor of axonal endings, spines and mitochondrial profiles. DSIP injection 1 hour before stress conditions result in appearance of flattened axonal endings stretched out on the spine head. Usually such contacts have two active zones set at angle to each other. The mcrease in synaptic vesicles number and total length of active zone suggests DSIPinduced stimulation of release apparatus work coupled with activation of additional post-synaptic receptors.
Previously we have reported that neocortex of hibematmg animals have highly developed protein synthetic stmctures and complexes of axe-axosornatic and axo-axodendritic contacts - so called “serial synapses”. It is likely that these peculiarities play an important role in adaptation process to low temperature. In ttns connection the present work aims to provide a compar~on studies of sensorimotor cortex of ground squirrels and rats duting normothetia and hypothermia (animals were cooled down to 25°C and were kept at this temperature during hvo hours). The lowering of temperature in rats entails disorganization of cell structures to a variable degree. Many Irregularly shaped lipofuscin granules and secondary lysosomes can be seen in neurons. The majority of dendritic spines are flattened. The reaction of ground squirrels neocorlex to hypothermia, contrastingly, is manifested in activation of receptor medIated exocytosis, lengthening of dendritic spine neck and activation of axe-axonal contacts. The results of our investigation suggests that hibernating and non-hibernating rodents use fundamentally different adaptation strategies while adjusting to low temperatures.
144
136
FUNCTIONAL AND STRUCTURAL ANALYSIS OF DSD-I-PG GLYCOSAMINOGLYCANS
MEMBERS
IDENTIFICATION
A. Clernenr, S. Nadanaka’, K. Masayama’. C. Ueoka*, K. Sugahara*. C. Mandl. and A. Falssner Dcparlmcnl for Neurobiology, University of Hcldclbcrg, 69120 Hcidclberg, FRG; *Department of Blochcmlstry, Kobc Pharmaceutical University, 658 Kobc, Japan
OF
REGC%-1 AND
OF A FAMILY
OF CELL
REGGIE-2,
SURFACE
TWO
PROTEINS
INFXSHANDMAMMALS.
S. Lommel, T. Schulte, M. Jung and C.A.O. Shmmer. Faculty of Biology, University of Konstanz, D-18434 Konstanz, Germany The moncclonal antibody M 802 recognizes a protein on the surface of ne&om retinal ganglion cells (RGCs) and RGC axon.5 tn the continuously growing retina of adult goldfish, which we named reppie. Reggie is re-expressed on all RGCs and RGC axons atIer optic nerve lesion and during axon regeneration suggesting a function of reggie for fiber tract repair. Two cDNAs, reggie-I and reggie-2, were identified in fish, and the lesioninduced upregulation of retie-1 and reggie-2 mRN4.s in RGCs was confirmed by in sifts hybridization. Moreover, we isolated partial rat cDNA clones for reggie-1 and reggie-2 by PCR based
The chondrmtin sulfalc protcoglycan DSD-I-PG has been identificd with tbc monoclonal antibody (mAb) 473HD. The mAb spccifitally rcac& with a structure contained in chondroitin sulfate C (CS C) which is degraded by chondroitinascs ABC and AU. but not by chondroitinase ACII. Thcsc results suppon Ihc mtcrprctation that the cpitope is composed of a dermatan (DS)/CS C hybrid (designated DSD-1 for dcmatan Sulfate dependent). DSD-I-PG IS cxprcssd by ghal precursor cells m a developmentally regulated fashion and Westcm blot analysis of mouse brain extracLr documcntcd maximal expression during early postnatal stages. When coated as substrate. DSD-I-ffi promotes neurite outgrowth of embryonic hippocampal neurons in virro. This suppoaive effccl is pamally blocked by mAb 473HD. which suggests functional rclevancc of the DSD-I glycosaminoglycan (Gag). Further structural analysis indicated that the 473HD-cpitopc requires the sulfation of the Gag. Thus, pharmacological agents. which block sulfation, abolished the epitopc in the DSD-l-PG expressing oligodcndroglial Oli-ncu precursor cell hne, as shown by immunoprecipitation and immunocytochemistry. This conclusion is consistent with competition ELlSAs using various CS of similar srructure, but distinct sulfarc composmon. In order 10 obtam more detailed information about the DSD-1 structure, CS C was digested with chondroitinase ACII. The resulting fragments were SL%Cfractionated by HPLC and tested by competition ELISA. Prchmmary data indicate that the epitopc is contained m a 16mcr fraction and two fractions of higher oligosaccharidcs. Supported by DFG (Fa 159/S-1.2.3)
cloning and obtained evidence for reggie expression in the rat CNS by in siti-hybridization. The characterization and spatiotemporal expression of reggie will be presented. Supported by DFG and BM!3F
xi