Study of changes in the rat choroid plexus after peripheral
nerve injury using fluorescent conjugated dextran

a 2015

Study of changes in the rat choroid plexus after peripheral nerve injury using fluorescent conjugated dextran

KUKLOVÁ, Adéla, Peter SOLÁR, Marek JOUKAL, Ilona KLUSÁKOVÁ, Petr DUBOVÝ et. al.

Základní údaje

Originální název

Study of changes in the rat choroid plexus after peripheral nerve injury using fluorescent conjugated dextran

Název česky

Studium změn plexus choroideus laboratorního potkana po poškození periferní nervu pomocí fluorescenčně značeného dextranu

Autoři

KUKLOVÁ, Adéla, Peter SOLÁR, Marek JOUKAL, Ilona KLUSÁKOVÁ a Petr DUBOVÝ

Vydání

Morphology 2015, 2015

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

30000 3. Medical and Health Sciences

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Impakt faktor

Impact factor: 0.924

Organizační jednotka

Lékařská fakulta

ISSN

Klíčová slova česky

Kolmerovy buňky, plexus choroideus, chronické poškození nervu, makrofágy

Klíčová slova anglicky

Kolmer cells, choroid plexus, chronic constriction injury, macrophages

Změněno: 16. 9. 2015 07:43, doc. MUDr. Marek Joukal, Ph.D.

Anotace

V originále

Aim. Peripheral nerve injury can lead to cellular and molecular changes in the peripheral and central nervous system structures which are not directly related with damaged nerve. We suppose that these structures can be influenced by damage associated molecular patterns (DAMPs) produced by Wallerian degeneration and diffused via blood-cerebrospinal fluid barrier (B-CSF-B). This barrier is present in the choroid plexus (CP) of brain ventricles. CP consists of highly vascularized stromal core with fenestrated capillaries and cuboidal cells covering the ventricular side. Epiplexal Kolmer cells (KC) attached on the surface of the cuboidal cells are involved in neuroimmunological regulations. The aim of our study was to investigate the changes of CP after peripheral nerve injury. For this purpose we used intravenously injected fluorescent conjugated dextran FluoroEmerald (FE) with different time of circulation. Methods. Twenty Wistar rats (male, 250-300g) were anaesthetized with a mixture of ketamine and xylazine and unilateral chronic constriction injury (CCI) of the sciatic nerve was performed. CCI operated rats were left to survive for 3 days (3D) (n=8) and 21 days (21D) (n=8). Four naive rats were used as control. After time of CCI survival the FE was intravenously applied after anesthesia to the left external jugular vein. Time of FE circulation was 5 hours (n=4) and 18 hours (n=4) in each period of CCI survival. Then, the rats were deeply anaesthetized, perfused transcardially by Zamboni´s fixative and the brains were removed and fixed for three days. Coronal cryostat sections (20 µm) were cut through the lateral and third ventricles. Distribution of FE and simultaneous immunohistochemical detection of resident (ED2) and activated (ED1) macrophages, antigen presenting cells (APC; MHC-II), T-cells (OX-52) and dendritic cells (OX-42) were analyzed using fluorescence microscope. Results. FE particles were found inside the cuboidal cells, the immune stromal cells and the KC. Statistically significant increase of number of FE+ cells was found in CP after 3 and 21 days of CCI and FE circulation for 18, but not for 5hours. Most of FE+ cells were found in CP 21 days from CCI and circulation for 18 hours. FE particles were detected inside stromal cells of the CP which displayed immunophenotypes of activated macrophages (ED1+) and antigen presenting cells (MHC-II+). KC without FE particles were immunostained for ED1, MHC-II and OX42. Number of OX42+ cells in CP increased with duration of nerve compression. FE positive KC were simultaneously positive for ED2 and OX52, respectively. Conclusion. CP responds to the chronic peripheral nerve damage with presence of FE+ cells in the stroma. Therefore, we assume that CP is a possible way how proinflammatory molecules from peripheral nerve injury could spread into the central nervous system. These CP changes occur with prolonged peripheral nerve damage by contrast to dorsal root ganglia. We also demonstrated increased penetration of immune cells through B-CSF-B induced by peripheral nerve injury. These changes could be caused by alteration in tight junctions.

Návaznosti

MUNI/A/1215/2014, interní kód MU

Název: Studium hemato-likvorové bariéry laboratorního potkana po poškození periferního nervu

Investor: Masarykova univerzita, Studium hemato-likvorové bariéry laboratorního potkana po poškození periferního nervu, DO R. 2020_Kategorie A - Specifický výzkum - Studentské výzkumné projekty

Citovat

KUKLOVÁ, Adéla, Peter SOLÁR, Marek JOUKAL, Ilona KLUSÁKOVÁ a Petr DUBOVÝ. Study of changes in the rat choroid plexus after peripheral nerve injury using fluorescent conjugated dextran. In Morphology 2015. 2015. ISSN 1213-8118.

@proceedings{1311177,
   author = {Kuklová, Adéla and Solár, Peter and Joukal, Marek and Klusáková, Ilona and Dubový, Petr},
   booktitle = {Morphology 2015},
   keywords = {Kolmer cells, choroid plexus, chronic constriction injury, macrophages},
   language = {eng},
   title = {Study of changes in the rat choroid plexus after peripheral nerve injury using fluorescent conjugated dextran},
   year = {2015}
}

TY  - CONF
ID  - 1311177
AU  - Kuklová, Adéla - Solár, Peter - Joukal, Marek - Klusáková, Ilona - Dubový, Petr
PY  - 2015
TI  - Study of changes in the rat choroid plexus after peripheral nerve injury using fluorescent conjugated dextran
KW  - Kolmer cells, choroid plexus, chronic constriction injury, macrophages
N2  - Aim. Peripheral nerve injury can lead to cellular and molecular changes in the peripheral and central nervous system structures which are not directly related with damaged nerve. We suppose that these structures can be influenced by damage associated molecular patterns (DAMPs) produced by Wallerian degeneration and diffused via blood-cerebrospinal fluid barrier (B-CSF-B). This barrier is present in the choroid plexus (CP) of brain ventricles. CP consists of highly vascularized stromal core with fenestrated capillaries and cuboidal cells covering the ventricular side. Epiplexal Kolmer cells (KC) attached on the surface of the cuboidal cells are involved in neuroimmunological regulations. The aim of our study was to investigate the changes of CP after peripheral nerve injury. For this purpose we used intravenously injected fluorescent conjugated dextran FluoroEmerald (FE) with different time of circulation. Methods. Twenty Wistar rats (male, 250-300g) were anaesthetized with a mixture of ketamine and xylazine and unilateral chronic constriction injury (CCI) of the sciatic nerve was performed. CCI operated rats were left to survive for 3 days (3D) (n=8) and 21 days (21D) (n=8). Four naive rats were used as control. After time of CCI survival the FE was intravenously applied after anesthesia to the left external jugular vein. Time of FE circulation was 5 hours (n=4) and 18 hours (n=4) in each period of CCI survival. Then, the rats were deeply anaesthetized, perfused transcardially by Zamboni´s fixative and the brains were removed and fixed for three days. Coronal cryostat sections (20 µm) were cut through the lateral and third ventricles. Distribution of FE and simultaneous immunohistochemical detection of resident (ED2) and activated (ED1) macrophages, antigen presenting cells (APC; MHC-II), T-cells (OX-52) and dendritic cells (OX-42) were analyzed using fluorescence microscope. Results. FE particles were found inside the cuboidal cells, the immune stromal cells and the KC. Statistically significant increase of number of FE+ cells was found in CP after 3 and 21 days of CCI and FE circulation for 18, but not for 5hours. Most of FE+ cells were found in CP 21 days from CCI and circulation for 18 hours. FE particles were detected inside stromal cells of the CP which displayed immunophenotypes of activated macrophages (ED1+) and antigen presenting cells (MHC-II+). KC without FE particles were immunostained for ED1, MHC-II and OX42. Number of OX42+ cells in CP increased with duration of nerve compression. FE positive KC were simultaneously positive for ED2 and OX52, respectively. Conclusion. CP responds to the chronic peripheral nerve damage with presence of FE+ cells in the stroma. Therefore, we assume that CP is a possible way how proinflammatory molecules from peripheral nerve injury could spread into the central nervous system. These CP changes occur with prolonged peripheral nerve damage by contrast to dorsal root ganglia. We also demonstrated increased penetration of immune cells through B-CSF-B induced by peripheral nerve injury. These changes could be caused by alteration in tight junctions.
ER  -

Podrobný výpis o publikaci