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Research Article
Hypoxia-ischemia alters distribution of lysosomal proteins in rat cortex and hippocampus
M. Troncoso, N. Bannoud, L. Carvelli, J. Asensio, A. Seltzer, M. A. Sosa
Biology Open 2018 7: bio036723 doi: 10.1242/bio.036723 Published 25 October 2018
M. Troncoso
1Laboratorio de Biología y Fisiología Celular “Dr. Franciso Bertini”, Instituto de Histología y Embriología – IHEM-CONICET-FCM-UNCuyo, 5500 Mendoza, Argentina
2Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Cuyo, 5500 Mendoza, Argentina
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N. Bannoud
1Laboratorio de Biología y Fisiología Celular “Dr. Franciso Bertini”, Instituto de Histología y Embriología – IHEM-CONICET-FCM-UNCuyo, 5500 Mendoza, Argentina
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L. Carvelli
1Laboratorio de Biología y Fisiología Celular “Dr. Franciso Bertini”, Instituto de Histología y Embriología – IHEM-CONICET-FCM-UNCuyo, 5500 Mendoza, Argentina
2Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Cuyo, 5500 Mendoza, Argentina
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J. Asensio
1Laboratorio de Biología y Fisiología Celular “Dr. Franciso Bertini”, Instituto de Histología y Embriología – IHEM-CONICET-FCM-UNCuyo, 5500 Mendoza, Argentina
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A. Seltzer
1Laboratorio de Biología y Fisiología Celular “Dr. Franciso Bertini”, Instituto de Histología y Embriología – IHEM-CONICET-FCM-UNCuyo, 5500 Mendoza, Argentina
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M. A. Sosa
1Laboratorio de Biología y Fisiología Celular “Dr. Franciso Bertini”, Instituto de Histología y Embriología – IHEM-CONICET-FCM-UNCuyo, 5500 Mendoza, Argentina
2Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Cuyo, 5500 Mendoza, Argentina
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  • ORCID record for M. A. Sosa
  • For correspondence: msosa@fcm.uncu.edu.ar
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    Fig. 1.

    Detection of astrogliosis employing an anti-GFAP antibody in normal and injured brains by HI. (A) Control Cx and HIP; (B) Ipsilateral (IL HI) Cx and HIP subjected to HI. A,B: 10× magnification. Image B shows the amplification of the area indicated by dashed lines (20× magnification). Scale bars: 150 μm.

  • Fig. 2.
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    Fig. 2.

    CatD expression and distribution in control and HI rat cerebral cortex. (A) Immunodetection of Cat D in membrane and cytosol. (B,C) Densitometric band quantification corresponding to intermediate and mature forms, respectively. Bars represent the mean of relative optical densities (R.O.D.±s.e.m.) from three independent experiments. *P<0.05, **P<0.01, as compared to controls. IL, ipsilateral; CL, contralateral brain hemisphere. β-tubulin was used as loading control.

  • Fig. 3.
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    Fig. 3.

    CatD expression and distribution in control and HI rat hippocampus. (A) Immunodetection of Cat D in membrane and soluble fractions. (B,C) Densitometric quantification of bands corresponding to intermediate and mature Cat D forms respectively. Bars represent the mean of relative optical densities (R.O.D.±s.e.m.) from three independent experiments. *P<0.05, **P<0.01, ***P<0.001, as compared to controls. IL, ipsilateral; CL, contralateral brain hemisphere. β-tubulin was used as loading control.

  • Fig. 4.
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    Fig. 4.

    PSAP expression and distribution in control and HI rat cerebral cortex. (A) Immunodetection of PSAP in membrane and cytosolic fractions. The same membrane from Fig. 2 was stripped as detailed in the Materials and Methods section and processed for PSAP or tubulin detection. (B) Densitometric quantification of the bands obtained in A. Bars represent the mean of relative optical densities (R.O.D.±s.e.m.) from three independent experiments.*P<0.05, **P<0.01, as compared to controls. IL, ipsilateral; CL, contralateral brain hemisphere. β-tubulin was used as loading control.

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    Fig. 5.

    PSAP expression and distribution in HI rat hippocampus. (A) Immunodetection of PSAP in membrane and cytosolic fractions. The same membrane from Fig. 3 was stripped as detailed in the Materials and Methods section and processed for PSAP or tubulin detection. (B) Densitometric quantification of the bands obtained in A. Bars represent the mean of relative optical densities (R.O.D.±s.e.m.) from three independent experiments. *P<0.05, **P<0.01, as compared to controls. IL, ipsilateral; CL, contralateral hemisphere. β-tubulin was used as loading control.

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    Fig. 6.

    Effect of HI on the integrity of LAMP-1. Immunodetection of LAMP-1 in membranes and cytosol of control and HI Cx. Ponceau Red protein staining was used as loading control. A representative image of two independent experiments is shown. IL, ipsilateral; CL, contralateral brain hemisphere; M, membranes; Cyt, Cytosol.

  • Fig. 7.
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    Fig. 7.

    Effect of HI on the integrity of LAMP-1 protein. Immunodetection of LAMP-1 in membranes and cytosol from control and HI hippocampus. Ponceau Red staining was used as loading control. A representative image of two independent experiments is shown. IL, ipsilateral; CL, contralateral brain hemisphere; M, membranes; Cyt, Cytosol.

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    Fig. 8.

    Distribution of αMan and βNAG enzyme activity. (A) In the cerebral cortex (Cx) and (B) in the hippocampus (HIP). Values are expressed as percentage (mean) of specific enzymatic activity (U/mg protein)±s.e.m. from three independent experiments. IL, ipsilateral; CL, contralateral brain hemisphere; αMan, α-mannosidase; βNAG, N-acetyl β-D-glucosaminidase.

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  • Hypoxia-ischemia
  • Excitotoxicity
  • Lysosomal enzymes
  • Lysosomes

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Research Article
Hypoxia-ischemia alters distribution of lysosomal proteins in rat cortex and hippocampus
M. Troncoso, N. Bannoud, L. Carvelli, J. Asensio, A. Seltzer, M. A. Sosa
Biology Open 2018 7: bio036723 doi: 10.1242/bio.036723 Published 25 October 2018
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Research Article
Hypoxia-ischemia alters distribution of lysosomal proteins in rat cortex and hippocampus
M. Troncoso, N. Bannoud, L. Carvelli, J. Asensio, A. Seltzer, M. A. Sosa
Biology Open 2018 7: bio036723 doi: 10.1242/bio.036723 Published 25 October 2018

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