PMID- 32321539
OWN - NLM
STAT- MEDLINE
DCOM- 20201130
LR  - 20201130
IS  - 2045-8118 (Electronic)
IS  - 2045-8118 (Linking)
VI  - 17
IP  - 1
DP  - 2020 Apr 22
TI  - Culture-induced changes in mRNA expression levels of efflux and SLC-transporters 
      in brain endothelial cells.
PG  - 32
LID - 10.1186/s12987-020-00193-5 [doi]
LID - 32
AB  - BACKGROUND: The complexity of the neurovascular unit (NVU) poses a challenge in 
      the investigations of drug transport across the blood-brain barrier (BBB) and the 
      function of the brain capillary endothelium. Several in vitro models of the brain 
      capillary endothelium have been developed. In vitro culture of primary 
      endothelial cells has, however, been reported to alter the expression levels of 
      various brain endothelial proteins. Only a limited number of studies have 
      addressed this in detail. The aim of the present study was to investigate mRNA 
      levels of selected BBB transporters and markers in in vitro models of the BBB 
      based on bovine primary endothelial cells and compare these to the levels 
      estimated in freshly isolated bovine brain capillaries. METHODS: Brain 
      capillaries were isolated from bovine cerebral cortex grey matter. Capillaries 
      were seeded in culture flasks and endothelial cells were obtained using a brief 
      trypsinization. They were seeded onto permeable supports and cultured in mono-, 
      non-contact- or contact co-culture with/without primary rat astrocytes. 
      mRNA-expression levels of the selected BBB markers and transporters were 
      evaluated using qPCR and monolayer integrity of resulting monolayers was 
      evaluated by measuring the transendothelial electrical resistance (TEER). 
      RESULTS: The capillary mRNA transcript profile indicated low expression of ABCC1 
      and CLDN1. The mRNA expression levels of TPA, OCLN, ABCB1, SLC2A1, SLC16A1 and 
      SLC7A5 were significantly decreased in all culture configurations compared to 
      freshly isolated bovine brain capillaries. ALP, VWF, ABCC1 and ABCC4 were 
      upregulated during culture, while the mRNA expression levels of F11R, TJP1, 
      CLDN5, CLDN1 and ABCG2 were found to be unaltered. The mRNA expression levels of 
      VWF, ALP, ABCB1 and ABCC1 were affected by the presence of rat astrocytes. 
      CONCLUSION: The endothelial mRNA transcript profile in bovine capillaries 
      obtained in this study correlated nicely with profiles reported in mice and 
      humans. Cultured endothelial cells drastically downregulated the mRNA expression 
      of the investigated SLC transporters but maintained expression of efflux 
      transporter and junctional protein mRNA, implying that the bovine in vitro BBB 
      models may serve well to investigate basic barrier biology and in vivo permeation 
      of passively permeating compounds and efflux transporter substrates but may be 
      less well suited for investigations of SLC-mediated transport.
FAU - Goldeman, C
AU  - Goldeman C
AD  - Department of Pharmacy, Faculty of Health and Medical Sciences, University of 
      Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark.
FAU - Ozgur, B
AU  - Ozgur B
AD  - Department of Pharmacy, Faculty of Health and Medical Sciences, University of 
      Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark.
FAU - Brodin, B
AU  - Brodin B
AD  - Department of Pharmacy, Faculty of Health and Medical Sciences, University of 
      Copenhagen, Universitetsparken 2, 2100, Copenhagen, Denmark. 
      Birger.brodin@sund.ku.dk.
LA  - eng
GR  - RIBBDD/Lundbeckfonden/
GR  - R155-2013-14113/Lundbeckfonden/
PT  - Journal Article
DEP - 20200422
PL  - England
TA  - Fluids Barriers CNS
JT  - Fluids and barriers of the CNS
JID - 101553157
RN  - 0 (Membrane Transport Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (Solute Carrier Proteins)
SB  - IM
MH  - Animals
MH  - Blood-Brain Barrier/*metabolism
MH  - Capillaries/*metabolism
MH  - Cattle
MH  - Cell Culture Techniques
MH  - Cells, Cultured
MH  - Cerebral Cortex/*blood supply
MH  - Coculture Techniques
MH  - Down-Regulation
MH  - Endothelial Cells/*metabolism
MH  - Endothelium, Vascular/*metabolism
MH  - Membrane Transport Proteins/*metabolism
MH  - RNA, Messenger/*metabolism
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Solute Carrier Proteins/metabolism
PMC - PMC7178567
OTO - NOTNLM
OT  - Blood-brain barrier
OT  - Brain endothelium
OT  - Co-culture
OT  - In vitro culture
OT  - SLC transporters
OT  - qPCR
COIS- The authors declare that they have no competing interests.
EDAT- 2020/04/24 06:00
MHDA- 2020/12/01 06:00
PMCR- 2020/04/22
CRDT- 2020/04/24 06:00
PHST- 2020/02/02 00:00 [received]
PHST- 2020/04/13 00:00 [accepted]
PHST- 2020/04/24 06:00 [entrez]
PHST- 2020/04/24 06:00 [pubmed]
PHST- 2020/12/01 06:00 [medline]
PHST- 2020/04/22 00:00 [pmc-release]
AID - 10.1186/s12987-020-00193-5 [pii]
AID - 193 [pii]
AID - 10.1186/s12987-020-00193-5 [doi]
PST - epublish
SO  - Fluids Barriers CNS. 2020 Apr 22;17(1):32. doi: 10.1186/s12987-020-00193-5.